Tricyclic compounds having sPLA2-inhibitory activities

ABSTRACT

The present invention provides a compound having sPLA 2  inhibiting activity. The compound represented by the formula (I):                    
     wherein R 1  is (a) C1 to C20 alkyl, C2 to C20 alkenyl, C2 to C20 alkynyl, carbocyclic groups, heterocyclic groups or the like; R 2  is CONH 2  or CONHNH 2 ; one of R 3  and R 4  is —(L 2 )-(acidic group) wherein L 2  is a group connecting with an acid group and the length of the connecting groups 1 to 5 atoms, and the other is a hydrogen atom; its prodrug, their pharmaceutically acceptable salt, or solvate thereof.

TECHNICAL FIELD

The present invention relates to a tricyclic compound effective forinhibiting sPLA₂-mediated fatty acid release.

BACKGROUND ART

sPLA₂ (secretory phospholipase A₂) is an enzyme that hydrolyzes membranephospholipids and has been considered to be a rate-determining enzymethat governs the so-called arachidonate cascade where arachidonic acid,the hydrolysis product, is the starting material. Moreover,lysophospholipids that are produced as by-products in the hydrolysis ofphospholipids have been known as important mediators in cardiovasculardiseases. Accordingly, in order to normalize excess functions of thearachidonate cascade and the lysophospholipids, it is important todevelop compounds which inhibit the liberation of sPLA₂-mediated fattyacids (for example, arachidonic acid), namely, compounds which inhibitthe activity or production of sPLA₂. Such compounds are useful forgeneral treatment of symptoms, which are induced and/or sustained by anexcess formation of sPLA₂, such as septic shock, adult respiratorydistress syndrome, pancreatitis, injury, bronchial asthma, allergicrhinitis, chronic rheumatism, arteriosclerosis, cerebral apoplexy,cerebral infarction, inflammatory colitis, psoriasis, cardiacinsufficiency, cardiac infarction, and so on. The participation of sPLA₂is considered to be extremely wide and, besides, its action is potent.

Examples of sPLA₂ inhibitors include compounds described in EP-620214(JP Laid-Open No. 010838/95, U.S. Pat. No. 5,578,634), EP-620215 (JPLaid-Open No. 025850/95, U.S. Pat. No. 5,684,034), EP-675110 (JPLaid-Open No. 285933/95, U.S. Pat. No. 5,654,326), WO 96/03120 (JPLaid-Open No. 505336/98), WO 96/03376 (JP Laid-Open No. 503208/98, U.S.Pat. No. 5,641,800), WO 96/03383 (JP Laid-Open No. 505584/98), WO97/21664 (EP-779271), WO 97/21716 (EP-779273), WO 98/18464 (EP839806),WO98/24437(EP846687), WO98/24756, WO98/24794, WO98/25609, WO99/51605,WO99/59999 and the like, or parabromophenacylbromide, mepacrine,manoaride, theilocien A and the like.

DISCLOSURE OF INVENTION

The object of the present invention is to provide tricyclic compoundshaving sPLA₂-inhibitory activities and being useful for treatment ofseptic shock, adult respiratory distress syndrome, pancreatitis, injury,bronchial asthma, allergic rhinitis, chronic rheumatism,arteriosclerosis, cerebral apoplexy, cerebral infarction, inflammatorycolitis, psoriasis, cardiac insufficiency, and cardiac infarction.

The present invention relates to I) a compound represented by theformula (I):

wherein R¹ is a group selected from (a) C1 to C20 alkyl, C2 to C20alkenyl, C2 to C20 alkynyl, carbocyclic groups, and heterocyclic groups,(b) the groups represented by (a) each substituted independently with atleast one group selected from non-interfering substituents, or (c)—(L¹)—R⁵ wherein L¹ is a divalent linking group of 1 to 18 atom(s)selected from hydrogen atom(s), nitrogen atom(s), carbon atom(s), oxygenatom(s), and sulfur atom(s), and R⁵ is a group selected from the groups(a) and (b);

one of R³ and R⁴ is —(L²)-(acidic group) wherein L² is an acid linkerhaving an acid linker length of 1 to 5 and the other is a hydrogen atom;

A ring is a group represented by the formula:

wherein R² is CONH₂ or CONHNH₂;

R¹⁸, R¹⁹, R²⁰, R²², and R²³ are each independently a hydrogen atom, orlower alkyl;

R²⁴ and R²⁵ are each independently a hydrogen atom, C1 to C6 alkyl,aryl, a halogen or aralkyl;

its prodrug, their pharmaceutically acceptable salt, or hydrate thereof.

In more detail, the present invention relates to II)-XIII).

II) A compound represented by the formula (II):

wherein R²⁴, R²⁵, and A ring are as defined above;

R⁶ is —(CH)_(m)—R⁹ wherein m is an integer from 1 to 6, and R⁹ is (d) agroup represented by the formula:

wherein a, c, e, n, q, t and v are each independently an integer from 0to 2; R¹⁰ and R¹¹ are each independently selected from a halogen, C1 toC10 alkyl, C1 to C10 alkyloxy, C1 to C10 alkylthio, optionallysubstituted phenyl, optionally substituted heteroaryl and C1 to C10haloalkyl; α is an oxygen atom or a sulfur atom; β is —CH₂— or —(CH₂)₂—;γ is an oxygen atom or a sulfur atom; b is an integer from 0 to 3, d isan integer from 0 to 4; f, p, and w are each independently an integerfrom 0 to 5; r is an integer from 0 to 7; and u is an integer from 0 to4, or R⁹ is (e) a member of (d) substituted with at least onesubstituent selected from the group consisting of C1 to C6 alkyl, C1 toC6 alkyloxy, C1 to C6 haloalkyloxy, C1 to C6 haloalkyl, phenyl, and ahalogen;

one of R⁷ and R⁸ is —(L³)—R¹² wherein L³ is represented by the formula:

wherein M is —CH₂—, —O—, —N(R⁵)—, or —S—; R¹³ and R¹⁴ are eachindependently a hydrogen atom, C1 to C10 alkyl, aryl, aralkyl, carboxy,or a halogen, and R¹⁵ is a hydrogen atom or C1 to C6 alkyl; and

R¹² is represented by the formula:

wherein R¹⁶ is hydrogen atom, a metal, or C1 to C10 alkyl; R¹⁷ isindependently a hydrogen atom or C1 to C10 alkyl; h is an integer from 1to 8;

its prodrug, their pharmaceutically acceptable salt, or hydrate thereof.

When the above b, d, f, p, r, u, and/or w are 2 or more, a plural numberof R¹⁰ or R¹¹ may be different from one another. When R¹⁰ is asubstituent on the naphthyl group, the substituent may substitute at anyarbitrary position on the naphthyl group. CH₂— and —(CH₂)₂— in β may besubstituted with R¹⁰.

III) A compound, its prodrug, their pharmaceutically acceptable salt, orhydrate thereof as described in I) or II), wherein said R¹ and R⁶ arerepresented by the formula:

wherein R¹⁰, R¹¹, b, d, f, p, r, u, w, α, β, and γ are as defined above.

When the above b, d, f, p, r, u, and/or w are 2 or more, a plural numberof R¹⁰ or R¹¹ may be different from one another. When R¹⁰ is asubstituent on the naphthyl group, the substituent may substitute at anyarbitrary position on the naphthyl group. —CH₂— and —(CH₂)₂— in β may besubstituted with R¹⁰.

IV) A compound, its prodrug, their pharmaceutically acceptable salt, orhydrate thereof as described in any one of I) to III), wherein said R¹and R⁶ are represented by the formula:

wherein R¹⁰, R¹¹, p, u, and w are as defined above.

When the above p, u, and/or w are 2 or more, a plural number of R¹⁰ orR¹¹ may be different from one another.

V) A compound, its prodrug, their pharmaceutically acceptable salt, orhydrate thereof as described in any one of I) to IV), wherein said R³and R⁷ are —O—(CH₂)_(m)—COOH (m is as defined above).

VI) A compound represented by the formula (III):

wherein R¹⁰, A ring, and m are as defined above,

its prodrug, their pharmaceutically acceptable salt, or hydrate thereof.

VII) A compound, its prodrug, their pharmaceutically acceptable salt, orhydrate thereof as described in any one of I) to VI), wherein said R² is—CONH₂.

VIII) A compound, its prodrug, their pharmaceutically acceptable salt,or hydrate thereof as described in any one of I) to VII), wherein saidR¹⁸, R¹⁹, R²⁰, R²¹, R²², and R²³ are hydrogen atoms.

IX) A pharmaceutical composition containing a compound as described inany one of I) to VIII) as an active ingredient.

X) A pharmaceutical composition as described in IX), which is forinhibiting sPLA₂.

XI) A pharmaceutical composition as described in IX), which is fortreatment or prevention of inflammatory diseases.

XII) Use of a compound of any one of I) to VII) for preparation of apharmaceutical composition for treating inflammatory diseases.

XIII) A method for treating a mammal, including a human, to alleviatethe pathological effects of inflammatory diseases, which comprisesadministration to said mammal of a compound as described in any one ofI) to VIII) in a pharmaceutically effective amount.

In the present specification, the term “alkyl” employed alone or incombination with other terms means a straight- or branched chainmonovalent hydrocarbon group having a specified number of carbon atoms.An example of the alkyl includes methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl,n-octyl, n-nonyl, n-decanyl, n-undecanyl, n-dodecanyl, n-tridecanyl,n-tetradecanyl, n-pentadecanyl, n-hexadecanyl, n-heptadecanyl,n-octadecanyl, n-nonadecanyl, n-eicosanyl and the like.

The term “alkenyl” employed alone or in combination with other terms inthe present specification means a straight- or branched chain monovalenthydrocarbon group having a specified number of carbon atoms and at leastone double bond. An example of the alkenyl includes vinyl, allyl,propenyl, crotonyl, isopentenyl, a variety of butenyl isomers and thelike.

The term “alkynyl” used in the present specification means a straight orbranched chain monovalent hydrocarbon group having a specified number ofcarbon atoms and at least one triple bond. The alkynyl may contain (a)double bond(s). An example of the alkynyl includes ethynyl, propynyl,6-heptynyl, 7-octynyl, 8-nonynyl and the like.

The term “carbocyclic group” used in the present specification means agroup derived from a saturated or unsaturated, substituted orunsubstituted 5 to 14 membered, preferably 5 to 10 membered, and morepreferably 5 to 7 membered organic nucleus whose ring forming atoms(other than hydrogen atoms) are solely carbon atoms. A group containingtwo to three of the carbocyclic group is also included in the abovestated group. An example of typical carbocyclic groups includes (f)cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and cyclooctyl); cycloalkenyl (such as cyclobutylenyl,cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl); phenyl,naphthyl, norbornyl, bicycloheptadienyl, indenyl, stilbenyl,terphenylyl, phenylcyclohexenyl, acenaphthyl, anthoryl, biphenylyl,bibenzylyl, and a phenylalkylphenyl derivative represented by theformula:

wherein x is an integer from 1 to 8.

The term “heterocyclic group” used in the present specification means agroup derived from monocyclic or polycyclic, saturated or unsaturatedheterocyclic nucleus having 5 to 14 ring atoms and containing 1 to 3hetero atoms selected from the group consisting of nitrogen atom, oxygenatom, and sulfur atom. An example of the heterocyclic group includespyridyl, pyrrolyl, furyl, benzofuryl, thienyl, benzothienyl, pyrazolyl,imidazolyl, phenylimidazolyl, triazolyl, isoxazolyl, oxazolyl,thiazolyl, thiadiazolyl, indolyl, carbazolyl, norharmanyl, azaindolyl,benzofuranyl, dibenzofuranyl, dibenzothiophenyl, indazolyl,imidazo[1,2-a]pyridinyl, benzotriazolyl, anthranilyl,1,2-benzisoxazolyl, benzoxazolyl, benzothiazolyl, purinyl, puridinyl,dipyridinyl, phenylpyridinyl, benzylpyridinyl, pyrimidinyl,phenylpyrimidinyl, pyrazinyl, 1,3,5-triazinyl, quinolyl, phthalazinyl,quinazolinyl, quinoxalinyl and the like.

Preferable are thienyl, furyl, thiazolyl, pyridyl as the heterocyclicring group in the R¹⁰ and R¹¹.

Preferred carbocyclic and heterocyclic groups in R¹ are (g) a grouprepresented by the formula:

wherein v is an integer from 0 to 2; R¹⁰ and R¹¹ are each independentlyselected from a halogen, C1 to C10 alkyl, C1 to C10 alkyloxy, C1 to C10alkylthio, optionally substituted phenyl, optionally substitutedheterocyclic group, and C1 to C10 haloalkyl, α is an oxygen atom or asulfur atom, β is —CH₂— or —(CH₂)₂—; γ is an oxygen atom or a sulfuratom; b is an integer from 0 to 3, d is an integer from 0 to 4; f, p,and w are an integer from 0 to 5; r is an integer from 0 to 7, and u isan integer from 0 to 4. When the above b, d, f, p, r, u, and/or w are 2or more, a plural number of R¹⁰ or R¹¹ may be different from oneanother. When R¹⁰ is a substituent on the naphthyl group, thesubstituent may be substituted at any arbitrary position on the naphthylgroup. —CH₂— and —(CH₂)₂— in β may be substituted with R¹⁰.

A more preferable example includes (h) a group represented by theformula:

wherein R¹⁰, R¹¹, α, β, and γ are the same as defined above, and y isindependently 0 or 1. When R¹⁰ is a substituent on the naphthyl group,the substituent may be substituted at any arbitrary position on thenaphthyl group. —CH₂— and —(CH₂)₂— in β may be substituted with R¹⁰.

The term “non-interfering substituent” in the present specificationmeans a group suitable for substitution of group (a) (e.g., “alkyl”,“alkenyl” “carbocyclic group” and “heterocyclic group”) in R¹ ontricyclic compound represented by the formula (I). An example of thenon-interfering substituents includes C1 to C10 alkyl, C2 to C6 alkenyl,C2 to C6 alkynyl, C7 to C12 aralkyl (such as benzyl and phenethyl), C7to C12 alkaryl, C3 to C8 cycloalkyl, C3 to C8 cycloalkenyl, phenyl,tolyl, xylyl, biphenylyl, C1 to C10 alkyloxy, C1 to C6 alkyloxy C1 to C6alkyl (such as methyloxymethyl, ethyloxymethyl, methyloxyethyl, andethyloxyethyl), C1 to C6 alkyloxy C1 to C6 alkyloxy (such asmethyloxymethyloxy and methyloxyethyloxy), C1 to C6 alkylcarbonyl (suchas methylcarbonyl and ethylcarbonyl), C1 to C6 alkylcarbonylamino (suchas methylcarbonylamino and ethylcarbonylamino), C1 to C6 alkyloxyamino(such as methyloxyamino and ethyloxyamino), C1 to C6alkyloxyaminocarbonyl (such as methyloxyaminocarbonyl andethyloxyaminocarbonyl), mono or di C1 to C6 alkylamino (such asmethylamino, ethylamino, dimethylamino, and ethylmethylamino), C1 to C10alkylthio, C1 to C6 alkylthiocarbonyl (such as methylthiocarbonyl andethylthiocarbonyl), C1 to C6 alkylsulfinyl (such as methylsulfinyl andethylsulfinyl), C1 to C6 alkylsulfonyl (such as methylsulfonyl andethylsulfonyl), C2 to C6 haloalkyloxy (such as 2-chloroethyloxy and2-bromoethyloxy), C1 to C6 haloalkylsulfonyl (such aschloromethylsulfonyl and bromomethylsulfonyl), C1 to C10 haloalkyl, C1to C6 hydroxyalkyl (such as hydroxymethyl and hydroxyethyl), C1-C6alkyloxycarbonyl (such as methyloxycarbonyl and ethyloxycarbonyl),—(CH₂)z—O—(C1 to C6 alkyl), benzyloxy, aryloxy (such as phenyloxy),arylthio (such as phenylthio), —(CONHSO₂R²⁰), formyl, amino, amidino,halogen, carbamyl, carboxyl, carbalkyloxy, —(CH₂)z—COOH (such ascarboxymethyl, carboxyethyl, and carboxypropyl), cyano, cyanoguanidino,guanidino, hydrazido, hydrazino, hydroxy, hydroxyamino, nitro,phosphono, —SO₃H, thioacetal, thiocarbonyl, carbonyl, carbocyclicgroups, heterocyclic groups and the like, wherein z is an integer from 1to 8 and R²⁰ is C1 to C6 alkyl or aryl.

Preferable are halogens, C1 to C6 alkyl, C1 to C6 alkyloxy, C1 to C6alkylthio, and C1 to C6 haloalkyl as the “non-interfering substituent”in the R¹. More preferable are halogens, C1 to C3 alkyl, C1 to C3alkyloxy, C1 to C3 alkylthio, and C1 to C3 haloalkyl.

The term “halogen” in the present specification means fluorine,chlorine, bromine, and iodine.

The term “cycloalkyl” in the present specification means a monovalentcyclic hydrocarbon group having a specified number of carbon atoms. Anexample of the cycloalkyl includes cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclooctyl and the like.

The term “cycloalkenyl” in the present specification means a monovalentcyclic hydrocarbon group having a specified number of carbon atoms andat least one double bond(s). An example of the cycloalkenyl includes1-cyclopropenyl, 2-cyclopropenyl, 1-cyclobutenyl, 2-cyclobutenyl and thelike.

In the present specification, an example of “alkyloxy” includesmethyloxy, ethyloxy, n-propyloxy, isopropyloxy, n-butyloxy, n-pentyloxy,n-hexyloxy and the like.

In the present specification, an example of “alkylthio” includesmethylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio,n-pentylthio, n-hexylthio and the like.

The term “acidic group” in the present specification means an organicgroup functioning as a proton donor capable of hydrogen bonding whenattached to a tricyclic nucleus through a suitable linking atom(hereinafter defined as “acid linker”). An example of the acidic groupincludes (k) a group represented by the formula:

wherein R¹⁶ is a hydrogen atom, a metal, or C1 to C10 alkyl; each R¹⁷ isindependently a hydrogen atom or C1 to C10 alkyl; h is an integer from 1to 8. Preferable is (1) —COOH, —SO₃H, or P(O)(OH)₂. More preferable is(m) —COOH. And preferable is also their ester and prodrug.

The term “acid linker” in the present specification means a divalentlinking group represented by a symbol —(L²)—, and it functions to jointricyclic nucleus to an “acidic group” in the general relationship. Anexample of it includes (n) a group represented by the formula:

wherein M is —CH₂—, —O—, —N(R¹⁵)—, or —S—, and R¹³ and R¹⁴ are eachindependently a hydrogen atom, C1 to C10 alkyl, aryl, aralkyl, carboxy,or halogens, wherein R¹⁵ is a hydrogen atom or C1-C6 alkyl. Preferableare (o) —O—CH₂—, —S—CH₂—, —N(R¹⁵)—CH₂—, —CH₂—CH₂—, —O—CH(CH₃)—, or—O—CH((CH₂)₂Ph)— wherein R¹⁵ is C1 to C6 alkyl and Ph is phenyl. Morepreferable is (p) —O—CH₂— or —S—CH₂—.

In the present specification, the term “acid linker length” means thenumber of atoms (except for hydrogen atoms) in the shortest chain of alinking group —(L²)— which connects tricyclic nucleus with the “acidicgroup”. The presence of a carbocyclic ring in —(L²)— counts as thenumber of atoms approximately equivalent to the calculated diameter ofthe carbocyclic ring. Thus, a benzene and cyclohexane ring in the acidlinker counts as two atoms in culculating the length of —(L²)—. Apreferable length is 2 to 3.

The term “haloalkyl” in the present specification means theaforementioned “alkyl” substituted with the aforementioned “halogen” atarbitrary position(s). An example of the haloalkyl includeschloromethyl, trifluoromethyl, 2-chloromethyl, 2-bromomethyl and thelike.

The term “hydroxyalkyl” in the present specification means theaforementioned “alkyl” substituted with hydroxy,at arbitraryposition(s). An example of the hydroxyalkyl includes hydroxymethyl,2-hydroxyethyl, 3-hydroxypropyl and the like. In this case,hydroxymethyl is preferable.

In the present specification, the term “haloalkyl” in “haloalkyloxy” isthe same as defined above. An example of it includes 2-chloroethyloxy,2-trifluoroethyloxy, 2-chloroethyloxy and the like.

The term “aryl” in the present specification means a monocyclic orcondensed cyclic aromatic hydrocarbon. An example of the aryl includesphenyl, 1-naphthyl, 2-naphthyl, anthryl and the like. Particularly,phenyl and 1-naphthyl are preferred.

The term “aralkyl” in the present specification means a group whereinthe aforementioned “alkyl” is substituted with the above-mentioned“aryl”. Such aryl may have a bond at any substitutable position. Anexample of it includes benzyl, phenethyl, phenylpropyl (such as3-phenylpropyl), naphthylmethyl (such as 1-naphtbylmethyl) and the like.

The term “alkyloxycarbonyl” in the present specification means C1-C6alkyloxycarbonyl. An example of the alkyloxycarbonyl includesmethyloxycarbonyl, ethyloxycarbonyl, n-propyloxycarbonyl and the like.

The term “acyl” in the present specification means C1-C6 alkylcarbonylor arylcarbonyl opptionally substituted with a halogen and the like. Anexample of the acyl includes acetyl, trifluoroacetyl, propionyl, benzoyland the like.

In the present specification, preferable are a halogen, C1-C10 alkyl,C1-C10 alkyloxy, C1-C10 alkylthio, and C1-C10 haloalkyl and the like assubstituents for “optionally substituted phenyl” and” “optionallysubstituted heterocyclic group”. These substituents may be substitutedwith one or more positions.

A group of preferable substituents as the R¹ to R³ of the compoundrepresented by the formula (I) will be shown in items (A) to (O).Preferable are hydrogen atoms as the R⁴, R¹⁸, R¹⁹, R²⁰, R²¹, R²², R²³,R²⁴, and R²⁵. Items (f) to (p) are the same group as described above.

As the R¹, (A): —(L¹)—R⁵, (B): —(CH₂)₁₋₂-(f), (C): —(CH₂)₁₋₂-(g), and(D): —(CH₂)₁₋₂-(h) are preferred.

As the R², (E): CONH₂ or CONHNH₂, and (F): CONH₂ are preferred.

As the R³, (G): -(n)-(k), (H): -(n)-(l), (I): -(n)-(m), (J): -(o)-(k),(K): -(o)-(l), (L): -(o)-(m), (M): -(p)-(k), (N): -(p)-(l), and (O):-(p)-(m) are preferred.

A preferred group of compounds represented by the formula (I) is shownbelow.

(R¹,R²,R³)=(A,E,G), (A,E,H), (A,E,I), (A,E,J), (A,E,K), (A,E,L),(A,E,M), (A,E,N), (A,E,O), (A,F,G), (A,F,H), (A,F,I), (A,F,J), (A,F,K),(A,F,L), (A,F,M), (A,F,N), (A,F,O), (B,E,G), (B,E,H), (B,E,I), (B,E,J),(B,E,K), (B,E,L), (B,E,M), (B,E,N), (B,E,O), (B,F,G), (B,F,H), (B,F,I),(B,F,J), (B,F,K), (B,F,L), (B,F,M), (B,F,N), (B,F,O), (C,E,G), (C,E,H),(C,E,I), (C,E,J), (C,E,K), (C,E,L), (C,E,M), (C,E,N), (C,E,O), (C,F,G),(C,F,H), (C,F,I), (C,F,J), (C,F,K), (C,F,L), (C,F,M), (C,F,N), (C,F,O),(D,E,G), (D,E,H), (D,E,I), (D,E,J), (D,E,K), (D,E,L), (D,E,M), (D,E,N),(D,E,O), (D,F,G), (D,F,H), (D,F,I), (D,F,J), (D,F,K), (D,F,L), (D,F,M),(D,F,N), and (D,F,O).

BEST MODE FOR CARRYING OUT THE INVENTION

The compounds of the invention represented by the formula (I) can besynthesized in accordance with well-known method described in chemicaljournals. The compounds of the invention represented by the formula (I)can also be synthesized in accordance with the following methods A to E.Although representative methods are exemplified, enlarged rings can alsobe synthesized in a similar manner.

wherein R¹, R³, R⁴, R⁵, R¹⁸, R¹⁹, R²⁰, R²¹, R²⁴, and R²⁵ are as definedabove, R²⁶ is lower alkyl, R²⁷CO— is a precursor of R¹, R²⁸ isoptionally substituted aryl, Hal are independently a halogen.

(Process 1)

A mixture of the compound (IV) and the compound (V) is stirred at 40° C.to 90° C., preferably 50 to 70° C. for 3 to 36 h, preferably 12 to 24 hto give the quaternary salt. To a solution of the obtained quaternarysalt in a solvent such as 1,2-dichloroethane or acetonitrile is added abase such as 1,8-diazabicyclo[5.4.0]-7-undecene (DBU) or triethylamine,and the resulting mixture is stirred at 40° C. to 90° C., preferably 50to 70° C. for 3 to 36 h, preferably 12 to 24 h. After the reactionmixture is subjected to a usual work-up, the compound (VII) can beobtained.

The compound (IV) is commercial available or can be synthesized inaccordance with the method described in J. Med. Chem., 3636-58(1996).The compound (V) can be synthesized in accordance with the methoddescribed in Synth. Commun. 24, 2557 (1994).

(Process 2)

The present process is performed by Friedel-Crafts reaction. To asolution of the compound (VI) in a solvent such as 1,2-dichloroethane ordichloromethane are slowly added R²⁷COHal and a Lewis acid such asAlCl₃, SbF₅, or BF₃ at −78° C. to 10° C., preferably −20° C. to icebath, and the reaction mixture is stirred at −10° C. to 10° C.,preferably 0° C. to 10° C. for 5 to 30 min, preferably 10 to 20 min.This reaction can be performed without solvent by dissolving thecompound (VI) in R²⁷COHal and in accordance with the above-mentionedprocedure. After the reaction mixture is subjected to a usual work-up,the compound (VII) can be obtained (Ref: J. Med. Chem., 39,3636-58(1996)).

(Process 3)

The present process includes the reduction of an ester group of a sidechain at 2-position and a carbonyl group of 3-position at the same time.To a solution of the compound (VII) in a solvent such as tetrahydrofuranor dichloromethane are added a reducing agent (e.g., a mixed agent ofsodium borohydride and a Lewis acid such as aluminum chloride), and themixture is reacted at 20° C. to 100° C., preferably 20° C. to 50° C. for1 to 5 h, preferably 1 to 3 h to obtain the compound (VIII).

(Process 4)

To a solution of the compound (VIII) in a solvent such as1,2-dichloroethane or tetrahydrofuran are added Hal—C(═O)—C(═O)—Hal(e.g., oxalyl chloride) and a base such as N-methylmorpholine ortriethylamine, and the mixture is stirred at 30 to 70° C., preferably 40to 60° C. for 1 to 10 h, preferably, 3 to 6 h. The reaction mixture ispoured into a cold aqueous ammonium solution and the resulting mixtureis stirred for 5 to 30 min, preferably, 10 to 20 min. After the reactionmixture is subjected to a usual work-up, the compound (IX) can beobtained.

(Process 5)

The present process includes the conversion of a hydroxy group tohalogen. To a solution of the compound (IX) is added triphenylphosphineand N-bromosuccinimide, and the mixture is reacted at 0° C. to 50° C.,preferably 0° C. to 20° C. for 1 to 10 h, preferably for 1 to 5 h toobtain the compound (X). It can be synthesized by using phosphoroustribroimde in accordance with the method described in Org. Synth Coll.Vol. 2, p-358, or by using triphenylphosphine and bromine in accordancewith the method described in J. Am. Chem. Soc., 107, 5238 (1995).

(Process 6)

The present process includes the preparation of the phosphonium salt. Amixture of the compound (X) and triphenylphosphine in a solvent such asacetonitrile or toluene is reacted at 80 to 150° C., preferably 100 to120° C. for 5 to 72 h, preferably 10 to 24 h to obtain the compound(XI).

(Process 7)

The present process is for constructing a ring by Wittig reaction. To asolution of the compound (XI) in a solvent such as acetonitrile, ortetrahydrofuran is added a base such as1,8-dizabicyclo[5.4.0]-7-undecene (DBU), potassium t-butoxide, and themixture is reacted at 20° C. to 120° C., preferably 80° C. to 100° C.for 3 to 24 h, preferably 5 to 10 h to obtain the compound (XII).

(Process 8)

The present process includes the reduction of the double bond byhydrogenation. To a solution of the compound (XII) in a solvent such astetrahydrofuran, methanol or ethyl acetate is added a catalyst such asPalladium-Carbon, and the mixture is reacted under hydrogen atmosphereat room temperature for 1 to 5h, preferably 1 to 2 h to yield thecompound (XIII).

wherein R¹, R³, R⁴, R⁵, R¹⁸, R¹⁹, R²⁴, R²⁵, R²⁶, R²⁷, and Hal are asdefined above. R²⁹ is a methanesulfonyl group, p-toluenesulfonyl or thelike.

(Process 1)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 1.

(Process 2)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 2.

(Process 3)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 3.

(Process 4)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 4.

(Process 5)

The present process is for the conversion of a hydroxy group at2-position to a good leaving group followed by the reduction of acarbonyl group of a side chain at 1-position. A solution of the compound(XVIII) in a solvent such as dichloromethane or the like is reacted withR²⁹—Cl in the presence of a base such as triethylamine or the like at 0°C. to 80° C., preferably 0° C. to 20° C. for 1 to 5 h, preferably 1 to 2h. A solution of the obtained compound in a solvent such astetrahydrofuran is reacted with a reductant such as sodium borohydrideat 0° C. to 80° C., preferably 0° C. to 20° C. for 1 to 5 h, preferably1 to 2 h to give the compound (XIX).

(Process 6)

The present step is for constructing a ring. To a solution of thecompound (XIX) in a solvent such as tetrahydrofuran, ordimethylformamide is added a base such as sodium hydride, or potassiumtert-butoxide at 0 to 100° C., preferably 20 to 50° C. for 1 to 8 h,preferably 1 to 3 h to give the compound (XX).

wherein R¹, R³, R⁴, R⁵, R¹⁸, R¹⁹R²⁰, R²¹, R²⁴, R²⁵, R²⁷, and Hal are asdefined above, one of R³⁰ and R³¹ is a hydrogen atom, the other is aprotecting group of an amino group.

(Process 1)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 1.

(Process 2)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 2.

(Process 3)

The present process is for the reduction of a carbonyl group at3-position methylene. To a solution of Lewis acid (e.g., AlCl₃ or thelike) in a solvent such as dichloromethane or tetrahydrofuran is slowlyadded a reducing agent such as borane-tert-butylamine complex or sodiumborohydride at −20° C. to 10° C., preferably in an ice bath, and thereaction mixture is stirred for 5 to 30 min, preferably 10 to 20 min. Tothe resulting reaction mixture is added a solution of the compound(XXIII) in a solvent such as dichloromethane or tetrahydrofuran at −20°C. to 10° C. preferably in an ice bath and the mixture is stirred for 20to 30 min, and then at 15° C. to 40° C., preferably 20 to 30° C. for 1to 5 h, preferably 2 to 3 h. After a usual work-up, the compound (XXIV)can be obtained (Ref: J. Med. Chem., 39, 3636-58 (1996)).

(Process 4)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 4.

(Process 5)

The present process is for the deprotection of the protecting group ofan amino group and the construction of the ring. To a solution of thecompound (XXV) in a solvent such as dichloromethane or the like is addedan acid such as trifluoroacetic acid and the mixture is reacted at 0° C.to 80° C., preferably 20° C. to 50° C. for 1 to 20 h, preferably 3 to 8h to give the compound (XXVI).

wherein R¹, R³, R⁴, R²⁴, R²⁵, R²⁷, and Hal are as defined above, R³² islower alkyl.

(Process 1)

The present process is for hydrolysis of ester. The compound (XXVII) canbe synthesized in accordance with the method described in J. Org. Chem.,36, 813 (1971). To a solution of the compound (XXVII) in a solvent suchas tetrahydrofuran, ethanol, or dimethylsulfoxide is added a base suchas sodium hydroxide, potassium hydroxide at 20 to 150° C., preferably 50to 150° C. for 1 to 10 h, preferably 3 to 5 h to obtain the compound(XXVIII).

(Process 2)

The present process is for the decarboxylation. A solution of thecompound (XXVIII) in solvent such as dimethyl sulfoxide ordimethylformamide is reacted at 50 to 200° C., preferably 100 to 150° C.for 1 to 5 h, preferably 1 to 2 h.

(Process 3)

The present process may be carried out in accordance with the sameprocedure as that of the Method C—Process 3.

(Process 4)

The present process is the reduction of carboxylic acid. Diborane,sodium borohydride-aluminum chloride, diisobutylaluminum hydride,lithium aluminum hydride and the like can be used as a reducing agent.In case of using lithium aluminum hydride, to a solution of the compound(XXX) in tetrahydrofuran or diethyl ether is added lithium aluminumhydride at 0° C. to 100° C., preferably 20° C. to 50° C. for 1 to 5 h,preferably 1 to 2 to give the compound (XXXI).

(Process 5)

The present process may be carried out in accordance with the sameprocedure as that of the Method A—Process 4.

(Process 6)

The present process is for the oxidation of a hydroxy group and theconstruction of the pyridazine ring.

The present process can be performed by usual oxidation and thefollowing four types of oxidation are preferable.

i) PCC Oxidation (To a solution of the compound (XXXII) in a solventsuch as dichloromethane or the like is added pyridinium chlorochromate(PCC) and the mixture is reacted at −20 to 60° C., preferably 0 to 40°C. for 1 to 30 h, preferably 3 to 20 h to yield the aimed oxidant.)(Ref:Tetrahedron Lett., 2647-2650 (1975))

ii) Swern Oxidation (Dichloromethane is cooled at −78° C., and to thesolution are added successively oxalyl chloride, dimethyl sulfoxide, andthe compound (XXXII). The mixture is warmed to −45° C. to 0° C., andreacted for 1 to 30 h, preferably 1 to 10 h and then is subjected tousual work-up to give the desired compound.) (Ref: J. Org. Chem., 43,2480-2482 (1978))

iii) Dess-Martin Oxidation (The oxidation can be carried out in solutionsuch as tetrahydrofuran or the like by reacting Dess-Martin reagent indimethyl sulfoxide.) (Ref: J. Org. Chem., 48, 4155-4156 (1983))

iv) Oxidation with a halogeno oxoacid (The compound (XXXII) is reactedin the presence of 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) with anoxidizing agent such as a halogeno oxoacid or the like in accordancewith the method described in J. Org. Chem., 52, 2559-2562 (1987) to givethe product. 2,2,6,6-tetramethylpiperidine-1-oxyl,4-methoxy-2,2,6,6-tetramethylpiperidine-1-oxyl,4-acetylamino-2,2,6,6-tetramethylpiperidine-1-oxyl,4-benzoyloxy-2,2,6,6-tetramethylpiperidine-1-oxyl,4-cyano-2,2,6,6-tetramethylpiperidine-1-oxyl or like the can be used asTEMPO or the like. Sodium hypochlorite, sodium hypobromite, sodiumbromite, high test hypochlorite or the like can be used as a halogenooxoacid. Ethyl acetate, acetonitrile, and dichlormethane can be used asa solvent.

Ring closing reaction can be carried out by reacting the obtainedcompound with hydrazine hydrate (NH₂NH₂H₂O) in a solvent such as ethanolor the like at 0° C. to 100° C., preferably 0° C. to 30° C. for 1 to 10h, preferably 2 to 3 h.)

wherein R¹, R³, R⁴, R²⁴, and R²⁵ are as defined above.

(Process 1)

The present process is for the preparation of N-alkoxyphthalimide byMitsunobu-reaction. The compound (XXXII) in a solvent such astetrahydrofuran or the like is reacted with triphenylphosphine,N-hydroxyphthalimide, diethyl azodicarboxylate, and the like at 0 to 80°C., preferably 10 to 30° C. for 1 to 5 h, preferably 1 to 2 h to obtainthe compound (XXXIV).

(Process 2)

The present process is for the ring formation. The compound (XXXIV) in asolvent such as dichloromethane or the like is reacted with reagentssuch as hydrazine, N-methylhydrazine or the like at 0° C. to 80° C.,preferably 10° C. to 30° C. for 1 to 5 h, preferably 1 to 2 h to givethe compound (XXXV).

Where a compound of the present invention has an acidic or basicfunctional group, a variety of salts having higher water solubility andmore physiologically suitable properties than those of the originalcompound can be formed. An example of typical pharmaceuticallyacceptable salts includes salts with alkali metal and alkaline earthmetal such as lithium, sodium, potassium, magnesium, aluminum and thelike, but it is to be noted that such pharmaceutically acceptable saltsare not limited thereto. A salt is easily manufactured from a free acidby either treating an acid in a solution with a base, or allowing anacid to be in contact with an ion exchange resin. Addition salts of thecompounds according to the present invention with relatively non-toxicinorganic bases and organic bases, for example, amine cation, ammonium,and quaternary ammonium derived from nitrogenous bases having a basicitysufficient for forming a salt of the compounds of the present inventionare included in the definition of “pharmaceutically acceptable salts”.(e.g., S. M. Berge et al., “Pharmaceutical Salts, “J. Phar. Sci., 66,1-19 (1977)). Furthermore, basic groups of a compound according to thepresent invention are reacted with a suitable organic or inorganic acidto form salts such as acetates, benzenesulfonates, benzoates,bicarbonates, bisulfates, bitartarate, borates, bromides, camcyrates,carbonates, chlorides, clubranates, citrates, edetates, edicirates,estrates, ethylates, fluorides, fumarates, gluseptates, gluconates,glutamates, glycolialsanyrates, hexylresorcinates, hydroxynaphthoates,iodides, isothionates, lactates, lactobionates, laurates, malates,malseates, manderates, mesylates, methylbromides, methylnitrates,methylsulfates, mucates, napcylates, nitrates, oleates, oxarates,palmitates, pantothenates, phosphates, polygalacturonates, salicirates,stearates, subacetates, sucinates, tanates, tartrates, tosylates,trifluoroacetates, trifluoromethanesulfonates, valerates and the like.In case of forming a hydrate, a questioned compound may be coordinatedwith a suitable number of water molecules.

In the case where a compound of the present invention has one or more ofchiral center(s), it may exist as an optically active member. Likewise,in the case where a compound contains alkenyl or alkenylene, there is apossibility of cis- and trans-isomers. Mixtures of R- and S-isomers aswell as of cis- and trans-isomers, and mixtures of R- and S-isomerscontaining racemic mixture are included in the scope of the presentinvention. Asymmetric carbon atom may exist also in a substituent suchas alkyl group. All such isomers are included in the present inventiontogether with these mixtures. In the case where a specified streoisomeris desired, either it is manufactured by applying a manner which hasbeen well known by those skilled in the art wherein a starting materialhaving an asymmetrical center which has been previously separated issubjected to stereospecific reaction to the starting material, or it ismanufactured by preparing a mixture of stereoisomers, and thereafterseparating the mixture in accordance with a well-known manner.

Prodrug is a derivative of the compound having a group which can bedecomposed chemically or metabolically, and such prodrug is a compoundaccording to the present invention which becomes pharmaceutically activeby means of solvolysis or by placing the compound in vivo under aphysiological condition. Although a derivative of the compoundsaccording to the present invention exhibits activity in both forms ofacid derivative and basic derivative, acid derivative is moreadvantageous in solubility, tissue affinity, and release control inmammal organism (Bungard, H., Design of Prodrugs, pp. 7-9, 21-24,Elsevier, Amsterdam, 1985). For instance, prodrugs each containing anacid derivative such as an ester which is prepared by reacting a basalacid compound with a suitable alcohol, or an amide which is prepared byreacting a basal acid compound with a suitable amine are well known bythose skilled in the art. Simple aliphatic or aromatic esters derivedfrom acid groups contained in the compounds according to the presentinvention are preferable prodrugs. More preferable is C1-C6 alkyl esterof acidic group (e.g., methyl ester, ethyl ester). Double ester such as(acyloxy)alkyl ester or ((alkyloxycarbonyl)oxy)alkyl ester type prodrugsmay be optionally manufactured.

The term “inhibit” means that release of fatty acid started by sPLA₂decreases significantly by the compounds:of the present invention fromviewpoint of prevention and treatment of disease. The term“pharmaceutically acceptable” means that carriers, diluents, oradditives are compatible with other ingredients in a formulation and arenot harmful for recipients.

The compounds of the present invention exhibit sPLA₂ inhibiting activityas per the description of the experimental examples which will bedescribed hereinafter. Accordingly, when a curatively effective amountof the compounds represented by the formulae (I), (II), and (III), theprodrug derivatives thereof, or their pharmaceutically acceptable salts,or their hydrate is administered to any of mammals (including humanbeing), it functions effectively as a curative medicine for diseases ofseptic shock, adult respiratory distress syndrome, pancreatitis, injury,bronchial asthma, allergic rhinitis, chronic rheumatism, arterialsclerosis, cerebral hemorrhage, cerebral infarction, inflammatorycolitis, psoriasis, cardiac failure, cardiac infarction.

The compounds of the present invention may be administered to a patientthrough a variety of routes including oral, aerosol, rectal,percutaneous, subcutaneous, intravenous, intramuscular, and nasalroutes. A formulation according to the present invention may bemanufactured by combining (for example, admixing) a curatively effectiveamount of a compound of the present invention with a pharmaceuticallyacceptable carrier or diluent. The formulation of the present inventionmay be manufactured with the use of well-known and easily availableingredients in accordance with a known method.

In case of manufacturing a composition according to the presentinvention, either active ingredients are admixed with a carrier, or theyare diluted with a carrier, or they are contained in a carrier in theform of capsule, sacheier, paper, or another container. In case offunctioning a carrier as a diluent, the carrier is a solid, semi-solid,or liquid material which functions as a medium. Accordingly, aformulation according to the present invention may be produced in theform of tablet, pill, powder medicine, intraoral medicine, elixir agent,suspending agent, emulsifier, dissolving agent, syrup agent, aerosolagent (solid in liquid medium), and ointment. Such a formulation maycontain up to 10% of an active compound. It is preferred to prepare acompound according to the present invention prior to administration.

Any suitable carrier which has been well known by those skilled in theart may be used for the formulation. In such formulation, a carrier isin the form of solid, liquid, or a mixture of solid and liquid. Forinstance, a compound of the present invention is dissolved into 4%dextrose/0.5% sodium citrate aqueous solution so as to be 2 mg/mlconcentration for intravenous injection. Solid formulation includespowder, tablet, and capsule. Solid carrier consists of one or more ofmaterial(s) for serving also as fragrant, lubricant, dissolving agent,suspension, binder, tablet disintegrator, capsule. A tablet for oraladministration contains a suitable excipient such as calcium carbonate,sodium carbonate, lactose, calcium phosphate and the like together witha disintegrator such as corn starch, alginic acid and the like and/or abinder such as gelatin, acacia and the like, and a lubricant such asmagnesium stearate, stearic acid, talc and the like.

In a powder medicine, a carrier is a finely pulverized solid which isblended with finely pulverized active ingredients. In a tablet, activeingredients are admixed with a carrier having required binding power ina suitable ratio, and it is solidified in a desired shape and size.Powder medicine and tablet contain about 1 to about 99% by weight of theactive ingredients being novel compounds according to the presentinvention. An example of suitable solid carriers includes magnesiumcarbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin,starch, gelatin, tragacanth gum, methyl cellulose, sodiumcarboxymethylcellulose, low-melting wax, and cocoa butter.

An axenic liquid formulation contains suspending agent, emulsifier,syrup agent, and elixir agent. Active ingredients may be dissolved orsuspended into a pharmaceutically acceptable carrier such as sterilewater, a sterile organic solvent, a mixture thereof and the like. Activeingredients may be dissolved frequently into a suitable organic solventsuch as propylene glycol aqueous solution. When finely pulverized activeingredients are dispersed into aqueous starch, sodiumcarboxylmethylcellulose solution, or suitable oil, the othercompositions can be prepared.

The dosage varies with the conditions of the disease, administrationroute, age and body weight of patient. In the case of oraladministration, the dosage can generally be between 0.01 to 50 mg/kg/dayfor adult.

The following examples are provided to further illustrate the presentinvention and are not to be constructed as limiting the scope thereof.

Abbreviations described below are used in the following examples.

Me:methyl

Et:ethyl

Ph:phenyl

Bn:benzyl

DBU:1,8-diazabicyclo[5.4.0]-7-undecene

EXAMPLE Example 1

Example 1

Process 1

A mixture of the compound (1) 8.90 g (44 mmol) and ethyl5-bromo-4-oxopentanate 10.20 g (45 mmol) was stirred at 60° C. for 1 h.After the reaction mixture was diluted with toluene, DBU 7.6 ml (50mmol) was added and the resulting mixture was stirred vigorously at 60°C. for 1 h. The reaction mixture was washed with water, then the toluenelayer was subjected to silica gel column chromatography to give thecompound (2) 7.90 g as colorless crystal.(yield: 55%) m.p.:81-84° C.

¹H-NMR(CDCl₃) δ 1.25 (3H, t, J=7.2 Hz), 2.66 (2H, t, J=10.8 Hz), 3.00(2H, t, J=10.8 Hz), 4.13 (2H, q, J=7.2 Hz), 5.16 (2H, s), 6.00 (1H, d,J=7.5 Hz), 6.31 (1H, t, J=7.5 Hz), 6.45 (1H, s), 7.12 (1H, s), 7.2-7.6(6H, m).

Example 1

Process 2

To a solution of the compound (2) 1.00 g (3.1 mmol) in toluene (10 ml)was added benzoyl chloride 0.55 ml (4.7 mmol) at room temperature andthe mixture was stirred at room temperature for 30 min and then at 80°C. for 1 h. The reaction mixture was washed with a saturated sodiumbicarbonate solution and dried. The toluene solution was absorbed bysilica gel and then the fraction eluted with toluene was recrystallizedwith ether to obtain the compound (3) 1.29 g colorless crystal. (yield:98%) m.p.: 102-103° C.

¹H-NMR(CDCl₃) δ 1.15 (3H, t, J=7.2 Hz), 2.44 (2H, t, J=5.7 Hz), 2.58(2H, t, J=5.7 Hz), 4.02 (2H, q, J=7.2 Hz), 5.21 (2H, s), 6.49 (1H, t,J=7.5 Hz), 6.59 (1H, s), 6.69 (1H, t, J=7.5 Hz), 7.3-7.6 (8H, m), 7.63(2H, d, J=7.8 Hz), 9.18 (1H, d, J=6.9 Hz).

Example 1

Process 3

To a solution of aluminum chloride 1.87 g (14.0 mmol) in tetrahydrofuran(30 ml) was added sodium borohydride 2.73 g (72.0 mmol) in an ice bath,and the mixture was stirred at same temperature for 10 min. To it wasadded the compound (3) 3.00 g (7.0 mmol) and the mixture was stirred atroom temperature for 2 h, and at 40° C. for 20 min. Then to the mixturewas added ether (100 ml), and 2N aqueous sodium hydroxide solution (50ml), and the resulting mixture was stirred vigorously. The ether layerwas washed with saturated aqueous sodium chloride solution, dried, andevaporated to give the compound (4) 2.61 g as colorless oil. (yield:100%) ¹H-NMR(CDCl₃) δ 1.93 (2H, m), 2.39 (2H, t, J=7.29 Hz), 3.68 (2H,t, J=6.0 Hz), 4.25 (2H, s), 5.17 (2H, s), 6.02 (1H, d, J=7.4 Hz), 6.27(1H, t, J=7.2 Hz), 6.59 (1H, s), 7.03 (2H, d, J=7.2 Hz), 7.20 (3H, m),4.40 (4H, m), 7.42 (2H, d, J=7.2 Hz).

Example 1

Process 4

To a solution of the compound (4) 2.60 g (7.0 mmol) in tetrahydrofuran(30 ml) was added oxalyl chloride (5.0 ml) and the mixture was stirredat the same temperature for 1 h. To the reaction mixture was added 28%aqueous ammonia solution at −20° C. for 20 min, and the mixture wasstirred at room temperature for 20 min. It was diluted with water andwas extracted with ethyl acetate. The extracts were washed with water,dried, and evaporated. The residue was recrystallized fromdichloromethane-ethanol to obtain the compound (5) 2.41 g as yellowcrystal. (yield: 78%) m.p.: 171-173° C.

¹H-NMR(CDCl₃-CD₃OD) δ 1.84 (2H, m), 2.95 (2H, t, J=7.5 Hz), 3.59 (2H, t,J=6.3 Hz), 4.31 (2H, s), 5.25 (2H, s), 6.46 (1H, d, J=7.2 Hz), 6.55 (1H,dd, J=6.9, 7.0 Hz), 7.10-7.50 (11H, m).

Example 1

Process 5

A suspension of the compound (5) 1.51 g (3.41 mmol) and 10%palladium-carbon (200 mg) in tetrahydrofuran was stirred under hydrogenatmosphere. After filtration of the catalyst and evaporation of thefiltrate, the residue was recrystallized from ethyl acetate to give thecompound (6) 1.09 g. (yield: 91%) m.p.: 167-169° C.

¹H-NMR(CDCl₃-CD₃OD) δ 1.85 (2H, m), 2.92 (2H, m), 3.61 (2H, t, J=6.3Hz), 4.25 (2H, s), 6.73 (1H, d, J=7.89 Hz), 6.82 (1H, t, J=7.8 Hz), 7.06(2H, d, J=6.9 Hz), 7.30 (4H, m).

Example 1

Process 6

To a solution of the compound (6) 480 mg (1.36 mmol) in DMF (5 ml) wereadded methyl bromoacetate 0.41 ml (4.34 mmol), potassium iodide 100 mg,and powder potassium hydroxide 114 mg (2.04 mmol) at room temperature,and the mixture was stirred for 30 min. To it were added methylbromoacetate 0.41 ml (4.34 mmol) and powder potassium hydroxide 114 mg(2.04 mmol), and the mixture was stirred for 30 min. The reactionmixture was poured into ethyl acetate, then the extract was washed withwater, dried, and evaporated, and the residue was absorbed on silicagel, then the fraction eluted with ethyl acetate was recrystallized fromethyl acetate to give the compound (7) 350 mg. (yield: 60%) m.p.:165-166° C.

¹H-NMR(CDCl₃) δ 1.85 (2H, m), 2.98 (2H, t, J=6.6 Hz), 3.40 (1H, brs),3.59 (2H, t, J=5.4 Hz), 3.79 (3H, s), 4.26 (2H, s), 4.74 (2H, s), 5.75(1H, brs), 6.31 (1H, t, J=7.8 Hz), 6.55 (1H, t, J=7.2 Hz), 6.65 (1H,brs), 7.04 (2H, d, J=6.6 Hz), 7.30 (3H, m), 7.34 (1H, d, J=6.9 Hz).

Example 1

Process 7

To a solution of the compound (7) 550 mg (1.30 mmol) in dichloromethane(7 ml) were added triphenylphosphine 407 mg (1.56 mmol) and NBS 278 mg(1.56 mmol) in an ice bath and the mixture was stirred at the sametemperature for 1 h. The reaction mixture was absorbed on silica gel,the fraction eluted with chloroform/ethyl acetate=1/1 was recrystallizedfrom ethyl acetate to give the compound (8) 520 mg. (yield: 83%) m.p.:175-176° C.

¹H-NMR(CDCl₃) δ 2.20 (2H, m), 3.01 (2H, t, J=7.2 Hz), 3.46 (2H, t, J=6.3Hz), 3.78 (3H, s), 4.29 (2H, s), 4.73 (2H, s), 5.70 (1H, brs), 6.28 (1H,d, J=7.8 Hz) 6.53 (1H, t, J=7.2), 6.80 (1H, brs), 7.07 (2H, d, J=6.6Hz), 7.25 (3H, m), 7.36 (1H, d, J=7.2 Hz).

Example 1

Process 8

A solution of the compound (8) 510 mg (1.05 mmol) and triphenylphosphine600 mg (2.25 mmol) in acetonitrile was refluxed for 6 h. The crystallineresidue which was obtained by evaporation under reduced pressure wasrecrystallized from ethyl acetate to give the compound (9) 695 mg.(yield: 89%) decomp.p.:225-227° C.

¹H-NMR(CDCl₃) δ 1.95 (2H, m), 3.23 (2H, t, J=6.8 Hz), 3.66 (2H, m), 3.78(3H, s), 4.41 (2H, s), 4.77 (2H, s), 5.85 (1H, brs), 6.28 (1H, d, J=7.8Hz), 6.55 (1H, t, J=7.2 Hz), 6.83 (1H, brs), 7.03 (2H, m), 7.13 (3H, m),7.40 (1H, d, J=7.2 Hz), 7.65 (15H, m).

Example 1

Process 9

To a suspension of the compound (9) 690 mg (0.92 mmol) in acetonitrile(10 ml) was added DBU 0.5 ml, and the mixture was stirred vigrously atroom temperature for 20 h. After evaporation under reduced pressure, tothe mixture were added ethyl acetate and 2N hydrochloric acid, and theresulting mixture was stirred. The ethyl acetate layer was washed withwater, dried, evaporated, then the residue was absorbed on silica geland fraction eluted with ethyl acetate was recrystallized to give thecompound (I-1) 186 mg as yellow crystal. (yield: 52%) m.p.: 173-175° C.

¹H-NMR(CDCl₃) δ 2.39 (2H, m), 2.71 (2H, t, J=7.4 Hz), 3.79 (3H, s), 4.20(2H, s), 4.70 (2H, s), 5.88 (1H, d, J=7.2 Hz), 6.25 (1H, t, J=7.2 Hz),6.44 (1H, t, J=5.7 Hz), 7.09 (2H, d, J=7.2 Hz), 7.15 (4H, m).

Elementary Analysis (for C₂₃H₂₂N₂O₄) Calcd.:C, 70.75; H, 5.68; N, 7.17.Found:C, 70.53; H, 5.68; N, 7.11.

Example 2

Example 2

Process 1

To a solution of the compound (I-1) 20 mg (0.05 mmol) in tetrahydrofuran(0.5 ml) was added 1N aqueous sodium hydroxide solution (0.2 ml), andthe mixture was stirred at room temperature for 10 min. After theprecipitated crystal was filtrated, washed with tetrahydrofuran, anddried, a solution of the crystal in water was stirred and adjusted with1N hydrochloric acid to pH=4.0. The precipitated crystal was filtrated,washed with water, and then dried to give the compound (I-2) 15 mg.(yield: 78%) decomp.p.: 238-242° C.

¹H-NMR(CDCl₃-CD₃OD) δ 2.42 (2H, m), 2.74 (2H, t, J=7.2 Hz), 4.22 (2H,s), 4.70 (2H, s), 5.99 (1H, d, J=7.5 Hz), 6.30 (1H, t, J=7.2 Hz), 6.33(1H, t, J=4.8 Hz), 7.10 (2H, d, J=6.6 Hz), 7.10-7.40 (4H, m).

Example 3

Example 3

Process 1

A suspension of t he compound (I-2) 10 mg (0.03 mmol) and 10%palladium-carbon (10 mg) in tetrahydrofuran (2 ml)-methanol (2 ml) wasstirred under hydrogen atmosphere. After filtration of the catalyst andevaporation of the filtrate, the precipitated crystal was filtrated togive the compound (I-3) 7 mg. (yield: 70%) decomp.p: 240-245° C.

¹H-NMR(CDCl₃-CD₃OD) d 2.00 (3H, m), 2.25 (1H, m), 2.73 (1H, m), 2.86(1H, m), 4.19 (2H, s), 4.31 (1H, m), 4.64(1H, s), 5.89 (1H, d, J=7.2Hz), 6.28 (1H, t, J=6.9 Hz), 7.07 (2H, d, J=6.9 Hz), 7.10-7.30 (4H, m).

Example 4

Example 4

Process 1

The compound (11) was synthesized in accordance with the methoddescribed in the reference (J. Org. Chem., 61, 2624 (1996)). yield: 65%.m.p.: 164.5-165.5° C.

¹H-NMR(CDCl₃) δ 2.52 (3H, S), 5.08 (2H, s), 6.76 (1H, dd, J=7.2, 3.6Hz), 6.86 (1H, d, J=3.3 Hz), 7.33-7.46 (5H, m), 8.16 (1H, d, J=7.2 Hz).

Example 4

Process 2

To a solution of the compound (11) 19.05 g (88.25 mmol) in chloroform130 ml was added a solution of phosphorus trichloride 18.4 ml (211 mmol)in chloroform 10 ml in an ice bath, and the mixture was stirred at roomtemperature for 30 min. The reaction mixture was poured into ice water,to the mixture was added gradually sodium hydroxide to be basified andthe resulting mixture was extracted with chloroform. The chloroformlayer was washed with brine, dried over magnesium sulfate andevaporated. The residue was dried under reduced pressure to give thecompound (12) 17.83 g. (yield: 100%).

¹H-NMR(CDCl₃) δ 2.53 (3H, s), 5.10 (2H, s), 6.72 (1H, dd, J=5.7, 2.4Hz), 6.76 (1H, d, J=2.4 Hz), 7.32-7.44 (5H, m), 8.32 (1H, d, J=6 Hz).

Example 4

Process 3

The compound (13) 6.68 g was obtained from the compound (12) 14.73 g ina manner similar to that described in Example 1—Process 1. (yield: 29%)m.p.: 84-87° C.

¹H-NMR(CDCl₃) δ 1.25 (3H, t, J=7.2 Hz), 2.64 (2H, t, J=7.7 Hz), 2.96(2H, t, J=7.7 Hz), 4.14 (2H, q, J=7.2 Hz), 5.00 (2H, s), 6.04 (1H, s),6.24 (1H, dd, J=7.2, 2.4 Hz), 6.59 (1H, d, J=2.4 Hz), 6.96 (1H, s),7.29-7.46 (5H, m), 7.67 (1H, d, J=7.5 Hz).

Example 4

Process 4

To a solution of the compound (13) 6.66 g (20.59 mmol) in toluene 60 mlwas added N-methylmorpholine 3.39 ml (30.83 mmol) and benzoyl chloride3.58 ml (30.84 mmol) in an ice bath and the mixture was stirred at roomtemperature for 1.5 h. The reaction mixture was poured into water, andthe resulting mixture was extracted with ethyl acetate. The extractswere washed with saturated aqueous sodium chloride solution, dried overmagnesium sulfate and evaporated under reduced pressure. The residue wasrecrystallized from ethyl acetate-methanol-n-hexane to give the compound(14) 6.79 g as light yellow green needle crystal. (yield: 77%) m.p.:137-138° C.

¹H-NMR(CDCl₃) d 1.16 (3H, t, J=7.2 Hz), 2.34-2.42 (2H, m), 2.47-2.55(2H, m), 4.02 (2H, q, J=7.2 Hz), 5.121 (2H, s), 6.17 (1H, s), 6.63 (1H,dd, J=2.7, 7.8 Hz), 6.780 (1H, d, J=2.7 Hz), 7.32-7.53 (8H, m),7.56-7.62 (2H, m), 9.66 (1H, d, J=7.8 Hz).

Example 4

Process 5

The compound (15) 5.49 g was obtained from the compound (14) 6.16 g in amanner similar to that described in Example 1—Process 3. (yield: 100%).

¹H-NMR(CDCl₃) δ 1.91 (2H, quint, J=6.9 Hz), 2.76 (2H, t, J=7.2 Hz), 3.68(2H, t, J=5.7 Hz), 4.22 (2H, s), 5.00 (2H, s), 6.16 (1H, s), 6.19 (1H,dd, J=7.5, 2.7 Hz), 6.67 (1H, d, J=2.7 Hz), 7.03 (2H, d, J=7.2 Hz),7.10-7.47 (9H,m).

Example 4

Process 6

To a solution of the compound (15) 215 mg (0.579 mmol) intetrahydrofuran 3 ml were added N-methylmorpholine 0.134 ml (1.129 mmol)and methyl oxalyl chloride 0.112 ml (1.218 mmol) in an ice bath and themixture was stirred at the same temperature for 40 min. The reactionmixture was poured into water, and the resulting mixture was extractedwith ethyl acetate. The extracts were washed with saturated aqueoussodium chloride solution, dried over magnesium sulfate and evaporatedunder reduced pressure. The residue (292 mg) was purified with silicagel column chromatography (eluted with n-hexane/ethyl acetate=2/1 to1/1) to give the compound (16) 106 mg as yellow foam. (yield: 34%).

¹H-NMR(CDCl₃) δ 2.00-2.12 (2H, m), 2.93 (2H, t, J=7.5 Hz), 3.86 (3H, s),3.94 (3H, s), 4.21 (2H, s), 4.29 (2H, t, J=6.3 Hz), 5.13 (2H, s), 6.54(1H, dd, J=7.5, 2.7 Hz), 7.03 (2H, d, J=6.3 Hz), 7.17-7.47 (8H, m), 7.55(1H, d, J=7.5 Hz), 7.70 (1H,brs).

Example 4

Process 7

To a solution of the compound (16) 1.563 g (2.83 mmol) intetrahydrofuran 20 ml was added 28% aqueous ammonia 50 ml and thesolution was sealed and stirred at 70° C. for 6.5 h. The reactionmixture was poured into water, and the resulting mixture was extractedwith ethyl acetate. The extracts were washed with brine, dried overmagnesium sulfate and evaporated under reduced pressure. The residue(1.204 g) was purified with silica gel column chromatography (elutedwith ethyl acetate to ethyl acetate/methanol=19/1) and successiverecrystallization (ethyl acetate-ethyl ether) to give the compound (17)758 mg as yellow needle crystal. (yield: 61%). m.p.: 157-158° C.

¹H-NMR(CDCl₃) δ 1.87 (2H, quint, J=6.6 Hz), 3.08 (2H, t, J=7.5 Hz), 3.63(2H, t, J=5.7 Hz), 4.22 (2H, s), 5.12 (2H, s), 5.74 (1H, brs), 6.51 (1H,dd, J=7.5, 2.4 Hz), 6.87 (1H, brs), 7.05 (2H, d, J=6.6 Hz), 7.16-7.46(8H, m), 7.52 (1H, d, J=7.5 Hz), 7.88 (1H, d, J=2.7 Hz).

Example 4

Process 8

The compound (18) 539 mg was obtained from the compound (17) 869 mg in amanner similar to that described in Example 1—Process 5. (yield: 78%)m.p.: 185-190° C.

¹H-NMR(DMSO-d₆) δ 1.70 (2H, quint, J=7 Hz), 2.86 (2H, t, J=7.5 Hz),2.37-2.46 (2H), m), 4.25 (2H, s), 4.38 (1H, m), 6.52 (1H, dd, J=7.5, 2.7Hz), 7.03 (2H, d, J=6.6 Hz), 7.15-7.32 (3H, m), 7.42 (1H, d, J=2.1 Hz),7.49 (1H, s), 7.88 (1H, d, J=7.5 Hz), 7.96 (1H, s), 10.51 (1H, brs).

Example 4

Process 9

To a solution of the compound (18) 42 mg (0.119 mmol) inN,N-dimethylformamide 1 ml were added ethyl 4-bromobutyrate 34 μl (0.238mmol) and 60% sodium hydride 4.8 mg (0.120 mmol) in an ice bath and themixture was stirred at the same temperature for 1 h. Furthermore, to thereaction mixture were added ethyl 4-bromobutyrate 17 μl (0.119 mmol) and60% sodium hydride 2.5 mg (0.063 mmol) and the mixture was stirred atthe room temperature for 1 h. The reaction mixture was poured intowater, and the resulting mixture was extracted with ethyl acetate. Theextracts were washed with water and brine successively, dried overmagnesium sulfate and evaporated under reduced pressure. The residue (71mg) was purified with a precoated SiO₂ plate (Merck, 0.5 mm, 20*20 cm)to give the compound (19) 24 mg as light yellow crystal (43%). m.p.:155-157° C.

¹H-NMR(CDCl₃) δ 1.25 (3H, t, J=7.2 Hz), 1.87 (2H, quint, J=6.7 Hz), 2.13(2H, quint, J=6.7 Hz), 2.49 (2H, t, J=7.2 Hz), 3.08 (2H, t, J=7.2 Hz),3.63 (2H, t, J=5.7 Hz), 4.08 (2H, t, J=6.6 Hz), 4.14 (2H, q, J=7.2 Hz),4.13 (2H, s), 5.91 (1H, brs), 6.42 (1H, dd, J=7.5, 2.7 Hz), 6.95 (1H,brs).

Example 4

Process 10

The compound (20) 874 mg was obtained from the compound (19) 591 mg in amanner similar to that described in Example 1—Process 7.

¹H-NMR(CDCl₃) δ 1.25 (3H, t, J=7 Hz), 2.07-2.25 (4H, m), 2.49 (2H, t,J=7.2 Hz), 3.11 (2H, t, J=7.2 Hz), 2.45 (2H, t, J=6.6 Hz), 4.08 (2H, t,J=6.5 Hz), 4.14 (2H, q, J=7.2 Hz), 4.25 (2H, s), 5.84 (1H, brs), 6.42(1H, dd, J=7.2,2.4 Hz), 6.93 (1H, brs), 7.07 (2H, d, J=6.3 Hz),7.17-7.32 (3H, m), 7.78 (1H, d, J=2.4 Hz).

Example 4

Process 11

The compound (21) 622 mg was obtained from the compound (20) 874 mg in amanner similar to that described in Example 1—Process 8. (2 processyield from the compound (19): 62%).

¹H-NMR(CDCl₃) δ 1.25 (3H, t, J=7 Hz), 1.80-2.04 (2H, m), 2.12 (2H,quint, J=6.5 Hz), 2.48 (2H, t, J=7.2 Hz), 3.27 (2H, t, J=7.4 Hz),3.66-3.84 (2H, m), 4.06 (2H, t, J=6 Hz), 4.14 (2H, q, J=7.2 Hz), 4.31(2H, s), 6.24 (1H, brs), 6.42 (1H, dd, J=7.5, 2.4 Hz), 6.93-7.80 (23H,m).

Example 4

Process 12

The compound (I-4) 121 mg was obtained from the compound (21) 544 mg ina manner similar to that described in Example 1—Process 9. (yield: 41%)m.p.: 152-154° C.

¹H-NMR(CDCl₃) δ 1.25 (3H, t, J=7.2 Hz), 2.08 (2H, quint, J=6.7 Hz),2.40-2.48 (2H, m), 2.47 (2H, t, J=7.2 Hz), 2.76 (2H, t, J=7.2 Hz), 3.96(2H, t, J=6 Hz), 4.13 (2H, q, J=7.2 Hz), 4.17 (2H, s), 5.61 (1H, brs),5.91 (1H, brs), 6.10 (1H, dd, J=7.5, 2.7 Hz), 6.34 (1H, t, J=4.8 Hz),6.85 (1H, d, J=2.4 Hz), 7.08 (2H, d, J=6.6 Hz), 7.15-7.30 (3H, m), 7.38(1H, d, J=7.5 Hz).

Example 5

Example 5

Process 1

The compound (I-5) 42 mg was obtained from the compound (I-4) 54 mg in amanner similar to that described in Example 2—Process 1. (yield: 83%)m.p.: 180-183° C.

¹H-NMR(DMSO-d₆) δ 1.93 (2H, quint, J=6.9 Hz), 2.27-2.42 (4H, m), 2.72(2H, t, J=7.5 Hz), 3.85 (2H, t, J=6 Hz), 4.19 (2H, s), 6.03 (1H, t,J=4.8 Hz), 6.20 (1H, dd, J=7.8, 2.7 Hz), 6.91 (1H, d, J=2.4 Hz),7.08-7.30 (6H, m), 7.52 (1H, brs), 7.73 (1H, d, J=7.5 Hz), 12.14 (1H,brs).

Example 6

Example 6

Process 1

To a solution of the compound (5) 4.00 g (9.05 mmol) in dichloromethane50 ml were added triethylamine 2.50 ml (18.1 mmol) and methanesulfonylchloride 0:91 ml (11.8 mmol) in an ice bath, the mixture was stirred for30 min. The mixture was diluted with ethyl acetate, and then the organiclayer was washed with water, dried, and evaporated under reducedpressure. The residue was dissolved in a solvent mixed withtetrahydrofuran 50 ml and methanol 50 ml and then to the solution wasadded sodium borohydride 2.0 g at room temperature for 30 min. Thereaction mixture was poured into ice water, and the resulting mixturewas extracted with ethyl acetate. The extracts were washed with water,dried and evaporated under reduced pressure. The residue wasrecrystallized from ethyl acetate to give the compound (22) 3.62 g.(yield: 77%). m.p.: 140-141° C.

¹H-NMR(CDCl₃) δ 2.02 (2H, m), 2.85 (2H, m), 2.92 (3H, s), 3.68 (1H, d,J=7.5 Hz), 4.21 (3H, s), 4.23 (2H, s), 5.16 (1H, d, J=15.0 Hz), 5.18(1H, d, J=15.0 Hz), 5.24 (1H, brs), 5.73 (1H d, J=7.5 Hz), 6.03 (1H,brs), 6.13 (1H, d, J=7.4 Hz), 6.31 (1H, t, J=7.4 Hz), 7.01 (2H, d, J=7.4Hz), 7.10-7.50 (9H, m).

Example 6

Process 2

To a solution of the compound (22) 2.10 g (4.02 mmol) in a solvent mixedwith N,N-dimethylformamide 20 ml and tetrahydrofuran 10 ml was added 60%sodium hydride 300 mg (5.00 mmol) under sonication at room temperaturefor 30 min and the mixture was stirred for 10 min under the samecondition. The reaction mixture was poured into ethyl acetate-water, theethyl acetate layer was washed with water and dried. The residueobtained by evaporation of the solvent under reduced pressure wasabsorbed on silica gel and the fraction eluted with toluene-ethylacetate (4:1 to 1:2) was recrystallized to give the compound (23) 930mg. (yield: 54%) m.p.: 201-203° C.

¹H-NMR(CDCl₃) δ 1.75 (1H, m), 2.04 (1H, m), 2.95 (2H, m), 4.07 (2H, m),4.14 (1H, d, J=17.4 Hz), 4.26 (1H, d, J=17.4 Hz), 5.11 (2H, s), 5.45(1H, brs), 6.06 (1H, d, J=7.2 Hz), 6.13 (1H, brs), 6.26 (1H, t, J=7.2Hz), 6.35 (1H, s), 7.04 (2H, d, J=6.9 Hz), 7.10-7.50 (9H, m).

Example 6

Process 3

To a solution of the compound (23) 680 mg in ethyl acetate 80 ml wasadded 10% palladium-carbon 300 mg and the mixture was stirred underhydrogen atmosphere. After filtration of the catalyst and evaporation ofthe filtrate, the residue was recrystallized from toluene to give thecompound (24) 296 mg. (yield: 57%) m.p.: 180-185° C.

¹H-NMR(CDCl₃) δ 1.70 (1H, m), 2.23 (1H m), 2.65 (1H, m), 3.10 (1H, m),3.86 (1H, m), 4.12 (1H, m), 4.19 (2H, s), 5.90 (1H, brs), 5.98 (1H, s),6.22 (1H, d, J=7.0 Hz), 6.31 (1H, t, J=7.2 Hz), 7.03 (2H, d, J=7.2 Hz),7.10-7.30 (4H, m), 10.33 (1H, brs).

Example 6

Process 4

To a solution of the compound (24) 350 mg (1.08 mmol) inN,N-dimethylformamide 4 ml were added methyl bromoacetate 0.31 ml (3.27mmol), potassium iodide 100 mg (0.60 mmol), and powder potassiumhydroxide 63 mg (1.12 mmol), and the mixture was stirred at roomtemperature under sonication for 15 min. The reaction mixture was pouredinto water and the mixture was extracted with ethyl acetate. Theextracts were washed with water and dried. The residue obtained byevaporation of the solvent under reduced pressure was purified withsilica gel column chromatography (eluted with toluene-ethyl acetate (1:1to 0:1) and successive recrystallization (ethyl acetate) to give thecompound (I-6) 152 mg. (yield: 35%). m.p.: 182-183° C.

¹H-NMR(CDCl₃) δ 1.70 (1H, m), 2.10 (1H, m), 2.87 (1H, m), 3.15 (1H, m),3.83 (3H, s), 4.03 (1H, m), 4.14 (1H, d, J=17.4 Hz), 4.19 (1H, m), 4.26(1H, d, J=17.4 Hz), 4.67 (1H, d, J=15.6 Hz), 4.71 (1H, d, J=15.6 Hz),5.40 (1H, brs), 5.89 (1H, d, J=7.2 Hz), 6.24 (1H, t, J=7.2 Hz), 6.43(1H, s), 6.68 (1H, brs), 7.04 (2H, d, J=7.5 Hz), 7.10-7.30 (4H, m).

Example 7

Example 7

Process 1

To a solution of the compound (I-6) 50 mg (0.12 mmol) in tetrahydrofuran1 ml was added 1N aqueous sodium hydroxide solution 0.4 ml at roomtemperature, and the mixture was stirred for 20 min. After to themixture was added 2N hydrochloric acid 0.2 ml, the resulting mixture wasevaporated. To the residue was added water, the mixture was stirred, andthen the precipitated crystal was filtrated to give the compound (I-7)41 mg. (yield: 85%) m.p.: 175-179° C.

¹H-NMR(CDCl₃) δ 1.63 (1H, m), 2.22 (1H, m), 2.80 (1H, mn), 3.10 (1H, m),3.87 (1H, m), 4.10 (1H, m), 4.17 (1H, d, J=16.5 Hz), 4.23 (1H, d, J=16.5Hz), 4.60 (1H, d, J=14.0 Hz), 5.80 (1H, brs), 5.99 (1H, d, J=7.5 Hz),6.11 (1H, s), 6.30 (1H, t, J=7.5 Hz), 7.02 (2H, d, J=6.9 Hz), 7.10-7.50(5H, m).

Example 8

Example 8

Process 1

To a solution of the compound (25) 10.9 g in N,N-dimethylformamide wasadded 60% sodium hydride 5.20 g in an ice bath, the mixture was stirredat room temperature for 15 min, and then to the mixture was addedp-methoxybenzylbromide 22.1 g in an ice bath, the resulting mixture wasstirred at room temperature for 1 h. To the reaction mixture was addedwater and the resulting mixture was extracted with toluene. The extractswere dried over magnesium sulfate. The residue obtained by evaporationof the solvent under reduced pressure was subjected to the silica gel 60g column chromatography to be eluted with a solution mixed with 50%ethyl acetate-toluene. The obtained crude crystal 15.97 g wasrecrystallized from ether-hexane to give the compound (26) 14.14 g aswhite crystal. (yield: 62%) m.p.: 74-76° C.

¹-NMR(CDCl₃) δ 2.51 (3H, s), 3.83 (3H, s), 5.01 (2H, s), 6.93 (2H, d),7.02-7.15 (2H, m), 7.35 (2 H, d), 8.09 (1H, m).

Example 8

Process 2

A solution of the compound (26) 5.00 g andN-(4-bromo-3-oxobutyl)phthalimide 6.46 g in dichloromethane 15 ml washeated at 65° C. in an oil bath for 2 h evaporating dichloromethane. Tothe precipitate crystal were added dichloroethane 50 ml and DBU 3.98 gand the mixture was refluxed in an oil bath for 2 h. To the reactionmixture was added water and the resulting mixture was extracted withchloroform. The extracts were dried over magnesium sulfate. The residueobtained by evaporation of the solvent under reduced pressure wassubjected to the silica gel 70 g column chromatography to be eluted with3% acetonitrile-chloroform. To the obtained fraction was added isopropylether, the precipitate crystal was filtrate to obtain the compound (27)6.41 g as yellow crystal (yield 69%). m.p.: 193-195° C.

¹H-NMR(CDCl₃) δ 3.05 (2H, t), 3.83 (3H, s), 3.96 (2H, t), 5.07 (2H, s),6.01 (1H, d), 6.31 (1H, t), 6.51 (1H, brs), 6.93 (2H, d), 7.19 (1H,brs), 7.38 (2H, d), 7.51 (1H, d), 7.68-7.71 (2H, m), 7.82-7.85 (2H, m).

Example 8

Process 3

The mixture of the compound (27) 5.89 g and hydrazine hydrate 6.7 ml inethanol 100 ml-tetrahydrofuran 100 ml was refluxed in an oil bath for 3h. The precipitated crystal was filtrated and the filtrate wasevaporated under reduced pressure. To the residue was added water, andthe resulting mixture was extracted with chloroform. The extracts weredried over magnesium sulfate and the residue obtained by evaporation wassubjected to the alumina 50 g column chromatography. The fraction elutedwith 5% methanol-chloroform gave the compound (28) 4.20 g as colorlessoil.

¹H-NMR(CDCl₃) δ 2.78 (2H, t), 2.98 (2H, t), 3.83 (3H, s), 5.09 (2H, s),6.02 (1H, d), 6.32 (1H, t), 6.45 (1H, brs), 6.93 (2H, d), 7.14 (1H, s),7.40 (2H, d), 7.52 (1H, d).

Example 8

Process 4

To a solution of the compound (28) 4.20 g in tetrahydrofuran 50 ml wasadded di-tert-butyl dicarbonate 3.16 g, and the mixture was stirred atroom temperature for 1 h. Evaporation of the solvent under the reducedpressure and recrystallization by adding to the residue isopropyl etherobtained the compound (29) 5.03 g as white crystal. (yield: 92%) m.p.:132-133° C.

¹H-NMR(CDCl₃) δ 1.43 (9H, s), 2.83 (2H, t), 3.41 (2H, m), 3.83 (3H, s),4.60 (1H, brs), 5.09 (2H, s), 6.03 (1H, d), 6.34 (1H, t), 6.44 (1H,brs), 6.93 (2H, d), 7.12 (1H, brs), 7.40 (2H, d), 7.52 (1H, d).

Example 8

Process 5

A solution of the compound (29) 2.00 g, N-methylmorpholine and benzoylchloride in dichloroethane 50 ml was refluxed for 2 h in an oil bath. Tothe reaction mixture was added water, and the resulting mixture wasextracted with chloroform. The extracts were dried over magnesiumsulfate and the residue obtained by evaporation was subjected to thesilica gel 50 g column chromatography. The fraction 2.52 g eluted with2.5% acetonitrile-chloroform was recrystallized from acetone-isopropylether to give the compound (30) 2.18 g as yellow crystal. (yield: 86%)m.p.: 135-137° C.

¹H-NMR(CDCl₃) δ 2.45 (2H, t), 3.19 (2H, m), 3.84 (3H, s), 4.33 (1H,brs), 5.14 (2H, s), 6.50 (1H, d), 6.61 (1H, s), 6.69 (1H, t), 6.95 (2H,d), 7.40 (2H, d), 7.43-7.63 (5H, m), 9.12 (1H, d).

Example 8

Process 6

To a solution prepared from aluminum chloride 533 mg and sodiumborohydride 757 mg in tetrahydrofuran 30 ml was added a solution of thecompound (30) 2.00 g in tetrahydrofuran 20 ml in an ice bath, themixture was stirred at room temperature for 2.5 h. The reaction mixturewas poured into ice water, and the resulting mixture was extracted withether. The extracts were dried over magnesium sulfate and thenevaporated. The solution of the residue in chloroform was subjected tothe silica gel 50 g column chromatography. The fraction 1.49 g elutedwith chloroform was recrystallized from acetone-isopropyl ether to givethe compound (31) 1.30 g as white crystal. (yield: 67%) m.p.: 92-93° C.

¹H-NMR(CDCl₃) δ 1.40 (9H, s), 2.87 (2H, t), 3.40 (2H, m), 3.83 (3H, s),4.23 (2H, s), 4.60 (1H, brs), 5.10 (2H, s), 6.04 (1H, d), 6.29 (1H, t),6.55 (1H, brs), 6.94 (2H, d), 7.00-7.23 (6H, m), 7.41 (2H, d).

Example 8

Process 7

To a solution of the compound (31) 1.73 g, N-methylmorpholine 0.78 ml indichloromethane 20 ml was added oxalyl chloride 1.55 ml indichloromethane 20 ml at −16 to −18° C. and the mixture was stirred for20 min. The reaction mixture was poured into a concentrated aqueousammonia solution 15 ml, insoluble substances were filtrated and thefiltrate was extracted with chloroform. The extracts were dried overmagnesium sulfate, and subjected to the silica gel 20 g columnchromatography. The fraction eluted with 20% acetonitrile-chloroform wasrecrystallized from isopropyl ether to obtain the compound (32) 1.76 gas yellow crystal. (yield: 89%) m.p.: 210-212° C. (decomposed).

¹H-NMR(DMSO-d₆) δ 1.33 (9H, s), 2.90 (2H, t), 3.14 (2H, m), 3.73 (3H,s), 4.29 (2H, s), 5.17 (2H, s), 6.44 (1H, d), 6.60 (1H, t), 6.84 (1H,brs), 6.89 (2H, d), 7.09-7.28 (6H, m), 7.45 (2H, d), 7.57 (1H, d), 7.67(1H, brs).

Example 8

Process 8

A suspension of the compound (32) 1.55 g and 10% palladium-carbon 160 mgin a solution mixed with tetrahydrofuran 50 ml-methanol 50 ml wasstirred for 5 h under hydrogen atmosphere. After filtration of the Pd-Cand evaporation of the filtrate, the residue was subjected to the silicagel 12 g column chromatography. The fraction 1.14 g eluted with 20%acetonitrile-chloroform was recrystallized from acetone-isopropyl etherto obtain the compound (33) 0.823 g as yellow crystal. (yield: 68%)m.p.: 148-149° C.

¹H-NMR(DMSO-d₆) δ 1.35 (9H, s), 2.96 (2H, t), 3.16 (2H, m), 4.32 (2H,s), 6.68 (1H, d), 6.81 (1H, m), 6.95 (1H, t), 7.09-7.31 (5H, m), 7.65(1H, d), 7.78 (1H, brs), 8.33 (1H, brs), 13.28 (1H, s).

Example 8

Process 9

A mixture of the compound (33) 650 mg, methyl bromoacetate 0.28 ml,potassium iodide 50 mg, potassium carbonate 206 mg inN,N-dimethylformamide 10 ml was stirred at room temperature for 6.5 h.The reaction mixture was poured into water and the precipitated crystalwas filtrated. A solution of the crude product in chloroform wassubjected to the silica gel 15 g column chromatography. The fraction 705mg eluted with a solution mixed with 75% acetonitrile-chloroform wasrecrystallized from acetone-isopropyl ether to give the compound (34)665 mg as yellow crystal. (yield: 88%) m.p.: 201-202° C.

¹H-NMR(CDCl₃) δ 1.38 (9H, s), 3.04 (2H, t), 3.41 (2H, m), 3.79 (3H, s),4.26 (2H, s), 4.73 (2H, s), 5.19 (1H, brs), 5.50 (1H, brs), 6.29 (1H,d), 6.53 (1H, t), 6.80 (1H, brs), 7.05-7.28 (5H, m), 7.35 (1H, d).

Example 8

Process 10

To a solution of the compound (34) 250 mg in dichloromethane 10 ml wasadded trifluoroacetic acid 0.38 ml at room temperature. The solution wasrefluxed for 8 h in an oil bath. After to the reaction mixture wereadded ice and an aqueous ammonia solution to be basified, the resultingmixture was extracted with chloroform. After the extracts were driedover magnesium sulfate and subjected to the silica gel 5 g columnchromatography. The fraction 236 mg eluted with a solution mixed with10% methanol-chloroform was recrystallized from methanol-ethyl acetateto give the compound. (I-8) 165 mg as light yellow brown crystal.(yield: 86%) m.p.: 210-213° C. (decomposed).

¹H-NMR(DMSO-d₆) δ 2.65 (2H, t), 3.60 (2H, t), 3.70 (3H, s), 4.27 (2H,s), 4.77 (2H, s), 6.31 (1H, d), 6.57 (1H, t), 7.03 (1H, brs), 7.15-7.31(5H, m), 7.38 (1H, brs), 7.69 (1H, d).

Example 9

Example 9

Process 1

To a solution of the compound (I-8) 145 mg in methanol 15 ml was added asolution of sodium hydroxide 1.5 ml at room temperature, and the mixturewas stirred for 75 min. To the residue obtained by evaporation ofmethanol under reduced pressure was added water, the resulting mixturewas acidified with 1N hydrochloric acid 1.55 ml, and the precipitatedcrystal was filtrated. The crude crystal was recrystallized frommethanol-water to give the compound (I-9) 121 mg as yellow crystal.(yield: 79%) m.p.: 196-198° C.

¹H-NMR(DMSO-d₆) δ 2.74 (2H, t), 3.63 (2H, t), 4.29 (2H, s), 4.60 (2H,s), 6.41 (1H, d), 6.71 (1H, t), 7.17-7.30 (5H, m), 7.39 (1H, brs), 7.67(1H, brs), 7.75 (1H, d).

Example 10

Example 10

Process 1

To a solution of the compound (35) 6.05 g (13.6 mmol) in methanol 50 mlwas added an aqueous potassium hydroxide solution (85%, 4.5 g, 68 mmol)20 ml, the mixture was refluxed for 4.5 h. After evaporation ofmethanol, the residue was acidified with 2N hydrochloric acid, and theresulting mixture was extracted with ethyl acetate. The extracts weredried over magnesium sulfate and evaporated to obtain the compound (36)5.60 g as yellow powder. (yield: 99%).

¹H NMR (DMSO-d₆) δ 5.33(2H, s), 6.84(1H, d, J=7.2 Hz), 6.99(1H, t, J=7.2Hz), 7.33-7.72(10H), 8.66(1H, d, J=7.2 Hz).

Example 10

Process 2

To a solution of the compound (36) 5.52 g (13.3 mmol) inN,N-dimethylformamide 25 ml was added p-toluenesufonic acid 253 mg (1.33mmol). The mixture was stirred at 115° C. for 30 min, acidified with 2Nhydrochloric acid, and extracted with ethyl acetate. The extracts werewashed with brine, dried over sodium sulfate and then evaporated. Theresidue was purified with the silica gel column chromatography to givethe compound (37) 4.85 as yellow powder. (yield: 98%).

¹H NMR (DMSO-d₆) δ 5.34(2H, s), 6.77(1H, d, J=7.5 Hz), 6.94(1H, s),6.95(1H, t, J=7.5 Hz), 7.37-7.70(10H), 7.98(1H, t, J=7.5 Hz).

Example 10

Process 3

To a solution of the compound (37) 2.95 g (7.9 mmol) in tetrahydrofuran20 ml was added sodium borohydride 1.5 g (40 mmol) and aluminum chloride3,18 g (24 mmol) in an ice bath. The mixture was stirred at roomtemperature for 4 h and then refluxed for 3.5 h. To the mixture wasadded ice water, the resulting mixture was acidified with 2Nhydrochloric acid, and extracted with ethyl acetate. The extracts werewashed with brine, dried over sodium sulfate and then evaporated. Theresidue was purified with the silica gel column chromatography to givethe compound (38) 1.37 g as light green powder. (yield: 48%).

¹H NMR (DMSO-d₆) δ 4.67(2H, s), 5.22(2H, s), 6.28(1H, d, J=7.2 Hz),6.55(1H, t, J=7.2 Hz), 6.85(1H, s), 7.14-7.52(10H), 7.63(1H, t, J=7.2Hz).

Example 10

Process 4

To a solution of the compound (38) 1.37 g (3.8 mmol) in tetrahydrofuran20 ml was added lithium aluminum hydride 1.16 g (31 mmol) and themixture was stirred at 55° C. for 2 h. To the mixture was added icewater, the resulting mixture was acidified with 2N hydrochloric acid,and extracted with ethyl acetate. The extracts were dried over sodiumsulfate and then evaporated to give the green oily substance. To asolution of the residue in tetrahydrofuran 20 ml was added oxalylchloride 1.67 ml (19 mmol) in an ice bath. After stirring for 30 min, tothe mixture was added a 28% aqueous ammonia solution 20 ml. Theresulting mixture was stirred for further 1 h, and extracted with ethylacetate. The extracts were dried over sodium sulfate and thenevaporated. The residue was purified with the silica gel columnchromatography to give the compound (39) 1.29 g as yellow powder.(yield: 82%).

¹H NMR (DMSO-d₆) δ 4.39(2H, s), 4.73(2H, d, J=5.7 Hz), 4.85(1H, t, J=5.7Hz), 5.26(2H, s), 6.43(1H, d, J=7.2 Hz), 6.64(1H, t, J=7.2 Hz),7.16-7.52(11H), 7.67(1H, d, J=7.2 Hz), 7.76(1H, brs).

Example 10

Process 5

To a solution of the compound (39) 600 mg (1.45 mmol) in tetrahydrofuran20 ml was added 10% palladium-carbon 60 mg, the mixture was stirred atroom temperature for 2 h under hydrogen atmosphere. The catalyst wasfiltrated and the solvent was evaporated. To a solution of the residuein N,N-dimethylformamide 10 ml were added potassium carbonate 600 mg(4.3 mmol), bezhydryl bromoacetate 530 mg (1.74 mmol), potassium iodide25 mg (0.15 mmol), and the mixture was stirred at room temperature for 2h. To the mixture was added ice water, the resulting mixture wasacidified with 2N hydrochloric acid, and extracted with ethyl acetate.The extracts were washed with brine, dried over sodium sulfate and thenevaporated. The residue was purified with the silica gel columnchromatography to give the compound (40) 550 mg as yellow powder.(yield: 69%.).

¹H NMR (DMSO-d₆) δ 4.42(2H, s), 4.75(2H, d, J=5.7 Hz), 4.86(1H, t, J=5.7Hz), 5.02(2H, s), 6.47(1H, d, J=7.5 Hz), 6.66(1H, t, J=7.5 Hz), 6.83(1H,s), 7.23-7.31(16H), 7.65(1H, brs), 7.74(1H, d, J=7.5 Hz).

Example 10

Process 6

To a solution of the compound (40) 372 mg (0.68 mmol) in dichloromethane20 ml was added a solution of Dess-Martin reagent 316 mg (0.75 mmol) indichloromethane 15 ml, and the mixture was stirred at room temperaturefor 5 min. The reaction mixture was washed with a 5% aqueous sodiumhydrogen carbonate solution, dried over sodium sulfate and evaporated.To a solution of the residue in ethanol 20 ml was added hydrazinehydrate 50 mg (1.0 mmol), and the mixture was stirred at roomtemperature for 2 h. Evaporation of the solvent and purification withthe silica gel column chromatography gave the compound (41) 45 mg asyellow powder. (yield; 12%).

¹H NMR (DMSO-d₆) δ 4.82(2H, s), 5.22(2H, s), 6.65(1H, d, J=7.5 Hz),6.84(1H, s), 7.18-7.32(16H), 7.53(1H, brs), 7.87(1H, brs), 8.20(1H, d,J=7.5 Hz), 9.65(1H, s).

Example 10

Process 7

To a solution of the compound (41) 38 mg (0.07 mmol) in dichloromethane1 ml were added anisole 0.1 ml and trifluoroacetic acid 0.1 ml at 010°C. After the mixture was stirred at room temperature for 15 min, it wasevaporated and diluted with ether. The precipitated powder wasfiltrated, washed with ether and dried to give the compound (I-10) 26 mgas orange powder. (yield: 99%).

¹H NMR (DMSO-d₆) δ 4.93(2H, s), 5.05(2H, s), 7.20-7.33(6H), 7.62(1H, t,J=6.9 Hz), 8.28(1H, brs), 8.41(1H, brs), 8.50(1H, d, J=6.9 Hz),9.84(1H,s).

Example 11

Example 11

Process 1

To a solution of the compound (40) 330 mg (0.60 mmol) in tetrahydrofuran5 ml were added triphenylphosphine 174 mg (0.66 mmol),N-hydroxyphthalimide 103 mg (0.63 mmol), and diethyl azodicarboxylate115 mg (0.66 mmol) in an ice bath, and the mixture was stirred for 30min. After evaporation of the solvent, the residue was purified withsilica gel chromatography to give the compound (42) 363 mg as yellowpowder. (yield: 87%).

¹H NMR (DMSO-d₆) δ 4.59(2H, s), 5.01(2H, s), 5.53(2H, s), 6.46(1H, d,J=7.2 Hz), 6.69(1H, t, J=7.2 Hz), 6.83(1H, s), 7.15-7.34(16H), 7.59(1H,brs), 7.72(1H, d, J=7.2 Hz), 7.80-7.85(4H).

Example 11

Process 2

To a solution of the compound (42) 325 mg (0.47 mmol) in dichloromethane6 ml was added N-methylhydrazine 22 mg (0.47 mmol) in an ice bath, andthe mixture was stirred at room temperature for 30 min. Insolublesubstances were filtrate and washed with dichloromethane. To thefiltrate was added methanol 5 ml, and then the mixture was stirred atroom temperature for 18 h. After evaporation of the solvent, the residuewas diluted with ether, then the precipitated powder was filtrated,dried to give the compound (43) 193 mg as yellow powder. (yield: 74%).

¹H NMR (DMSO-d₆) δ 4.29(2H, s), 4.93(2H, s), 4.99(2H, s), 6.35(1h, d,J=7.2 Hz), 6.61(1H, t, J=7.2 Hz), 6.85(1H, s), 7.18-7.37(16H), 7.73(1H,brs), 7.78(1H, d, J=7.2 Hz).

Example 11

Process 3

To a solution of the compound (43) 50 mg (0.09 mmol) in dichloromethane1 ml were added anisole 0.1 ml and trifluoroacetic acid 0.3 ml at 0° C.After the mixture was stirred at room temperature for 15 min, it wasevaporated and diluted with ether. The precipitated powder wasfiltrated, washed with ether and dried to give the compound (I-11)16 mgas yellow powder. (yield: 47%).

¹H NMR (DMSO-d₆) δ 4.29(2H, s), 4.70(2H, s), 4.91(2H, s), 6.38(1H, d,J=7.2 Hz), 6.68(1H, t, J=7.2 Hz), 7.18-7.32(5H), 7.39(1H, brs), 7.78(1H,d, J=7.2 Hz), 7.82(1H, brs).

The compounds (I-12) to (I-15) were synthesized by the same reactionsdescribed in the Examples 1 to Example 11. The physical data were shownin Tables 1 to 2.

TABLE 1

Compound ¹H-NMR: δ CDCl₃ (R¹ = Me), No. R¹ m.p. (° C.) CDCl₃—CD₃OD (R¹ =H) I-12 Me 196-198 2.34-2.44(2H, m), 2.74(2H, t, J= 7.4Hz), 3.79(3H, s),4.24(2H, s), 4.71 (2H, s), 5.25-5.85(2H, brs), 5.90(1H, d, J=7.5Hz),6.27(1H, t, J=7.2Hz), 6.45(1H, t, J=5.1Hz), 7.16(2H, d, J= 8.4Hz),7.27-7.58(8H, m) I-13 H 224-227 2.30-2.54(2H, m), 2.68-2.88(2H, m),4.28(2H, brs), 4.71(2H, s), 6.02(1H, m), 6.25-6.45(2H, m), 7.18(2H, d,J=7.8Hz), 7.28-7.60(8H, m)

TABLE 2

Compound ¹H-NMR: δ CDCl₃ (R¹ = Me), No. R¹ m.p. (° C.) DMSO-d₆ (R¹ = H)I-14 Me 218-220 2.74(1H, m), 2.93(1H, m), 3.82(3H, s), 4.14(1H, d,J=15.0Hz), 4.20(1H, d, J=15.0Hz), 4.18(2H, m), 4.67(1H, d, J=15.9Hz),4.71(1H, d, J=15.6Hz), 5.52 (1H, brs), 5.86(1H, d, J=7.5Hz), 5.87 (1H,s), 6.29(1H, t, J=7.59Hz), 6.73 (1H, brs), 7.06(2H, d, J=7.3Hz), 7.20-7.40(4H, m) I-15 H 242-243 2.75(2H, m), 3.90(1H, m), 4.20(1H, m), (dec.)4.20(2H, s), 4.64(1H, d, J=16.8Hz), 4.67(1H, d, J=16.8Hz), 5.55(1H, s),5.98(1H, d, J=7.2Hz), 6.37(1H, t, J= 7.2Hz), 7.01(1H, brs),7.10-7.30(6H, m), 7.54(1H, d, J=7.2Hz), 13.11(1H, brs)

Test Example Inhibition Test of Human Secretory Phospholipase A₂Analytical Experiment

In order to identify and evaluate an inhibitor of recombinant humansecretory phospholipase A₂, the following chromogenic assay is utilized.The assay herein has been applied for high volume screening wherein 96well microtiterplate is used. A general explanation for such assay isdescribed in “Analysis of Human Synovial Fluid Phospholipase A₂ on ShortChain Phosphatidylcholine-Mixed Micelles: Development of aSpectrophotometric Assay Suitable for a Micortiterplate Reader”(Analytical Biochemistry, 204, pp 190-197, 1992 by Laure. J. Reynolds.Lori L. Hughes and Edward A. Dennis: the disclosure of which isincorporated herein for reference.

Reagents

Reaction Buffer

CaCl₂.6H₂O (2.19 g/L)

KCl (7.455 g/L)

Bovine Serum Albumin (fatty acid free) (1 g/L) (Sigma A-7030)

Tris-HCl (3.94 g/L)

pH 7.5 (adjusted with NaOH)

Enzyme Buffer

0.05 M-AcONa

0.2 M-NaCl

pH 4.5 (adjusted with acetic acid)

Enzyme Solution

1 mg of sPLA₂ is dissolved in 1 ml of an enzyme buffer. Thereafter, thesolution is maintained at 4° C.

In the assay, 5 μl of the solution is diluted with 1995 μl of thereaction buffer to be used.

DTNB

198 mg of 5,5′-dithiobis-2-benzoic acid (manufactured by Wako PureChemicals) is dissolved in 100 ml of H₂O

pH 7.5 (adjusted with NaOH)

Substrate Solution

100 mg of racemic1,2-bis(heptanoylthio)-1,2-dideoxy-sn-glycero-3-phospholylcholine isdissolved in 1 ml of chloroform.

Triton-X 100

624.9 mg of Triton-X 100 is dissolved in 100 ml of the reaction buffer.

Enzyme Reaction: for 1 plate of Microtiterplate

1) 0.106 ml of the substrate solution is put in a centrifugal tube, andnitrogen gas is jetted to remove the solvent. 0.54 ml of Triton-X 100 isadded thereto, the mixture is stirred, thereafter it is sonified in abath type sonification to dissolve. To the resulting product are added17.8 ml of the reaction buffer and 0.46 ml of DTNB, and 0.18 ml each ofthe admixture is poured to wells of the 96 well microtiterplate.

2) 10 μl of a test compound (or solvent blank) are added in accordancewith alignment of plates which has been previously set.

3) Incubation is effected at 40° C. for 15 minutes.

4) 20 μl of an enzyme solution (sPLA₂) which has been previously diluted(50 ng/well) are added to start reaction (40° C., 30 minutes).

5) Changes in absorbancy for 30 minutes are measured by a plate reader,and inhibition activity was calculated (OD: 405 nm).

6) IC₅₀ was determined by plotting log concentration with respect toinhibition values within 10% to 90% inhibiting range.

Results of the human secretory phospholipase A₂ inhibition test areshown in the following Table 3.

TABLE 3 Compound No. IC₅₀ (μM) Compound No IC₅₀ (μM) I-2 0.008 I-9  1.59I-3 0.021 I-10 39.9 I-4 0.175 I-11 0.031 I-5 0.049 I-12 0.125 I-6 7.50I-13 0.010 I-7 0.269 I-14 0.725 I-8 21.1 I-15 0.035

Formulation Example

It is to be noted that the following Formulation Examples 1 to 9 aremere illustration, but not intended to limit the scope of the invention.The term “active ingredient” means the compounds represented by theformula (I), the prodrugs thereof, their pharmaceutical acceptablesalts, or their solvates.

Formulation Example 1

Hard gelatin capsules are prepared using of the following ingredients:

Dose (mg/capsule) Active ingredient 250 Starch, dried 200 Magnesiumstearate 10 Total 460 mg

Formulation Example 2

A tablet is prepared using of the following ingredients:

Dose (mg/tablet) Active ingredient 250 Cellulose, microcrystals 400Silicon dioxide, fumed 10 Stearic acid 5 Total 665 mg

The components are blended and compressed to form tablets each weighing665 mg.

Formulation Example 3

An aerosol solution is prepared containing the following components:

Weight Active ingredient 0.25 Ethanol 25.75 Propellant 22(chlorodifluoromethane) 74.00 Total 100.00

The active compound is mixed with ethanol and the admixture added to aportion of the propellant 22, cooled to −30° C. and transferred tofilling device. The required amount is then fed to stainless steelcontainer and diluted with the reminder of the propellant. The valveunits are then fitted to the container.

Formulation Example 4

Tablets, each containing 60 mg of active ingredient, are made asfollows.

Active ingredient 60 mg Starch 45 mg Microcrystals cellulose 35 mgPolyvinylpyrrolidone (as 10% solution in water) 4 mg Sodiumcarboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1 mg Total150 mg

The active ingredient, starch, and cellulose are passed through a No. 45mesh U.S. sieve, and the mixed thoroughly. The aqueous solutioncontaining polyvinylpyrrolidone is mixed with the resultant powder, andthe admixture then is passed through a No. 14 mesh U.S. sieve. Thegranules so produced are dried at 50° C. and passed through a No. 18mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate,and talc, previously passed through No. 60 mesh U.S. sieve, are thenadded to the granules which, after mixing, are compressed on a tabletmachine to yield tablets each weighing 150 mg.

Formulation Example 5

Capsules, each containing 80 mg of active ingredient, are made asfollows:

Active ingredient 80 mg Starch 59 mg Microcrystals cellulose 59 mgMagnesium stearate 2 mg Total 200 mg

The active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 45 mesh U.S. sieve, and filled into hardgelatin capsules in 200 mg quantities.

Formulation Example 6

Suppository, each containing 225 mg of active ingredient, are made asfollows:

Active ingredient 225 mg Saturated fatty acid glycerides 2000 mg Total2225 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2 g capacity and allowed to cool.

Formulation Example 7

Suspensions, each containing 50 mg of active ingredient per 5 ml dose,are made as follows:

Active ingredient 50 mg Sodium carboxymethyl cellulose 50 mg Syrup 1.25ml Benzoic acid solution 0.10 ml Flavor q.v. Color q.v. Purified waterto total 5 ml

The active ingredient is passed through a No. 45 U.S. sieve, and mixedwith the sodium carboxymethyl cellulose and syrup to form a smoothpaste. The benzoic acid solution, flavor and color are diluted with aportion of the water and added, with stirring. Sufficient water is thenadded to produce the required volume.

Formulation Example 8

An intravenous formulation may be prepared as follows:

Active ingredient 100 mg Isotonic saline 1000 ml

The solution of the above ingredients generally is administeredintravenously to a subject at a rate of 1 ml per minute.

Formulation Example 9

Composition of lyophilized preparations (in 1 vial) is made as follows:

Active ingredient 127 mg Trisodium citrate dihydrate 36 mg Mannitol 180mg

The above materials are dissolved in water for injection such that theconcentration of Active ingredient is 10 mg/g. The primary freezing stepis done for 3 hours at −40° C., the heat treating step for 10 hours at−10° C., and the re-freezing step for 3 hours at −40° C. Then, theprimary drying step is performed for 60 hours at 0° C., 10 Pa and thesecondary drying step for 5 hours at 60° C., 4 Pa. Thus the lyophilizedpreparation is obtained.

INDUSTRIAL APPLICABILITY

The compounds according to the present invention have sPLA₂ inhibitingactivity, so that the compounds of the invention inhibits sPLA₂-mediatedfatty acid (such as arachidonic acid) release, whereby it is effectivefor treating inflammatory diseases and the like.

What is claimed is:
 1. A compound represented by the formula (I):

wherein R¹ is a group selected from (a) C1 to C20 alkyl, C2 to C20alkenyl, C2 to C20 alkynyl, carbocyclic groups, and heterocyclic groups,(b) the groups represented by (a) each substituted independently with atleast one group selected from non-interfering substituents, or (c)—(CH₂)_(m)—R⁵ wherein m is an integer from 1 to 6, and R⁵ is a groupselected from the groups (a) and (b); one of R³ and R⁴ is —(L²)-(acidicgroup) wherein L² is represented by the formula:

wherein M is —CH₂—, —O—, —N(R¹⁵)—, or —S—; R¹³ and R¹⁴ are eachindependently a hydrogen atom, C1 to C10 alkyl, aryl, aralkyl, carboxy,or halogens, wherein R¹⁵ is a hydrogen atom or C1 to C6 alkyl; andacidic group is represented by the formula:

wherein R¹⁶ is hydrogen atom, a metal, or C1 to C10 alkyl; R¹⁷ isindependently a hydrogen atom or C1 to C10 alkyl; h is an integer from 1to 8 and the other is a hydrogen atom, A ring is a group represented bythe formula:

wherein R² is CONH₂ or CONHNH₂; R¹⁸, R¹⁹, R²⁰, R²¹, R²², and R²³ areeach independently a hydrogen atom, or lower alkyl; R²⁴ and R²⁵ are eachindependently a hydrogen atom, C1 to C6 alkyl, aryl, a halogen oraralkyl; its prodrug, their pharmaceutically acceptable salt, or hydratethereof.
 2. A compound represented by the formula (II):

wherein R²⁴, R²⁵, and A ring are as defined above; R⁶ is —(CH₂)_(m)—R⁹wherein m is an integer from 1 to 6, and R⁹ is (d) a group representedby the formula:

wherein a, c, e, n, q, t and v are each independently an integer from 0to 2; R¹⁰ and R¹¹ are each independently selected from a halogen, C1 toC10 alkyl, C1 to C10 alkyloxy, C1 to C10 alkylthio, optionallysubstituted phenyl, optionally substituted heteroaryl and C1 to C10haloalkyl; α is an oxygen atom or a sulfur atom; β is —CH₂— or —(CH₂)₂—;γ is an oxygen atom or a sulfur atom; b is an integer from 0 to 3, d isan integer from 0 to 4; f, p, and w are each independently an integerfrom 0 to 5; r is an integer from 0 to 7; and u is an integer from 0 to4, or R⁹ is (e) a member of (d) substituted with at least onesubstituent selected from the group consisting of C1 to C6 alkyl, C1 toC6 alkyloxy, C1 to C6 haloalkyloxy, C1 to C6 haloalkyl, phenyl, and ahalogen; one of R⁷ and R⁸ is —(L³)—R¹² wherein L³ is represented by theformula:

wherein M is —CH₂—, —O—, —N(R¹⁵)—, or —S—; R¹³ and R¹⁴ are eachindependently a |hydrogen atom, C1 to C10 alkyl, aryl, aralkyl, carboxy,or a halogen, and R¹⁵ is a hydrogen atom or C1 to C6 alkyl; and R¹² isrepresented by the formula:

wherein R¹⁶ is hydrogen atom, a metal, or C1 to C10 alkyl; R¹⁷ isindependently a hydrogen atom or C1 to C10 alkyl; h is an integer from 1to 8; its prodrug, their pharmaceutically acceptable salt, or hydratethereof.
 3. A compound, its prodrug, their pharmaceutically acceptablesalt, or hydrate thereof as claimed in claim 1, wherein said R¹ isrepresented by the formula:

wherein R¹⁰ and R¹¹ are each independently selected from a halogen, C1to C10 alkyl, C1 to C10 alkyloxy, C1 to C10 alkylthio, optionallysubstituted phenyl, optionally substituted heteroaryl and C1 to C10haloalkyl, α is an oxygen atom or a sulfur atom, β is —CH₂— or —(CH₂)₂—;γ is an oxygen atom or a sulfur atom; b is an integer from 0 to 3, d isan integer from 0 to 4; f, p, and w are each independently an integerfrom 0 to 5; r is an integer from 0 to 7, and u is an integer from 0 to4.
 4. A compound, its prodrug, their pharmaceutically acceptable salt,or hydrate thereof as claimed in claim 1, wherein said R¹ is representedby the formula:

wherein R¹⁰, R¹¹, p, u, and w are as defined above.
 5. A compound, itsprodrug, their pharmaceutically acceptable salt, or hydrate thereof asclaimed in claim 1, wherein said R³ and R⁷ are —O—(CH₂)_(m)—COOH (m isas defined above).
 6. A compound represented by the formula (III):

wherein R¹⁰, A ring, and m are as defined above, its prodrug, theirpharmaceutically acceptable salt, or hydrate thereof.
 7. A compound, itsprodrug, their pharmaceutically acceptable salt, or hydrate thereof asclaimed in claim 1, wherein said R² is —CONH₂.
 8. A compound, itsprodrug, their pharmaceutically acceptable salt, or hydrate thereof asclaimed in claim 1, wherein said R¹⁸, R¹⁹, R²⁰, R²¹, R²², and R²³ arehydrogen atoms.
 9. A pharmaceutical composition containing a compound asclaimed in claim 1 as an active ingredient.
 10. A method for treating amammal, including a human, to alleviate the pathological effects of adisease mediated by sPLA₂, which comprises administering to said mammala compound as claimed in claim 1 in a pharmaceutically effective amountwherein the disease is selected from the group consisting of adultrespiratory syndrome, pancreatitis, bronchial asthma, allergic rhinitis,chronic rheumatism, arteriosclerosis, cerebral infarction and psoriasis.